Modulation of hsp47 expression

ABSTRACT

Provided herein are compositions, methods and kits for modulating expression of target genes, particularly heat shock protein 47 (hsp47). The compositions, methods and kits may include nucleic acid molecules (for example, short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA) or short hairpin RNA (shRNA)) that modulate a gene encoding hsp47, for example, the gene encoding human hsp47. The composition and methods disclosed herein may also be used in treating conditions and disorders associated with hsp47 such as liver fibrosis, pulmonary fibrosis, peritoneal fibrosis and kidney fibrosis.

RELATED PATENT APPLICATIONS

This application claims the benefit of U.S. Provisional Application Ser.Nos. 61/372,072, filed Aug. 9, 2010, 61/307,412, filed Feb. 23, 2010 and61/285,149, filed Dec. 9, 2009 each entitled “Modulation of HSP47Expression” and which are incorporated herein by reference in theirentirety and for all purposes.

SEQUENCE LISTING

The instant application contains a Sequence Listing which is entitled220-PCT1_ST25_(—)07Dec-10.txt, said ASCII copy, created on Dec. 7, 2010and is 533 kb in size, is hereby incorporated by reference in itsentirety.

FIELD OF THE INVENTION

Provided herein are compositions and methods for modulating expressionof hsp47.

BACKGROUND OF THE INVENTION

Sato, Y., et al. disclose the administration of vitamin A-coupledliposomes to deliver small interfering RNA (siRNA) against gp46, the rathomolog of human heat shock protein 47, to liver cirrhosis rat animalmodels. Sato, Y., et al., Nature Biotechnology, vol. 26(4), p. 431-442(2008).

Chen, J-J., et al. disclose transfecting human keloid samples withHSP47-shRNA (small hairpin RNA) to examine proliferation of keloidfibroblast cells. Chen, J-J., et al., British Journal of Dermatology,vol. 156, p. 1188-1195 (2007).

PCT Patent Publication No. WO 2006/068232 discloses an astrocytespecific drug carrier which includes a retinoid derivative and/or avitamin A analog.

SUMMARY OF THE INVENTION

Compositions, methods and kits for modulating expression of target genesare provided herein. In various aspects and embodiments, compositions,methods and kits provided herein modulate expression of heat shockprotein 47 (hsp47), also known as SERPINH1. The compositions, methodsand kits may involve use of nucleic acid molecules (for example, shortinterfering nucleic acid (siNA), short interfering RNA (siRNA),double-stranded RNA (dsRNA), micro-RNA (miRNA) or short hairpin RNA(shRNA)) that bind a nucleotide sequence (such as an mRNA sequence)encoding hsp47, for example, the mRNA coding sequence for human hsp47exemplified by SEQ ID NO:1. In certain preferred embodiments, thecompositions, methods and kits disclosed herein inhibit expression ofhsp47. For example, siNA molecules (e.g., RISC length dsNA molecules orDicer length dsNA molecules) are provided that reduce or inhibit hsp47expression. Also provided are compositions, methods and kits fortreating and/or preventing diseases, conditions or disorders associatedwith hsp47, such as liver fibrosis, cirrhosis, pulmonary fibrosisincluding lung fibrosis (including ILF), kidney fibrosis resulting fromany condition (e.g., CKD including ESRD), peritoneal fibrosis, chronichepatic damage, fibrillogenesis, fibrotic diseases in other organs,abnormal scarring (keloids) associated with all possible types of skininjury accidental and jatrogenic (operations); scleroderma;cardiofibrosis, failure of glaucoma filtering operation; and intestinaladhesions.

In one aspect, provided are nucleic acid molecules (e.g., siNAmolecules) in which (a) the nucleic acid molecule includes a sensestrand and an antisense strand; (b) each strand of the nucleic acidmolecule is independently 15 to 49 nucleotides in length; (c) a 15 to 49nucleotide sequence of the antisense strand is complementary to asequence of an mRNA encoding human hsp47 (e.g., SEQ ID NO: 1); and (d) a15 to 49 nucleotide sequence of the sense strand is complementary to thea sequence of the antisense strand and includes a 15 to 49 nucleotidesequence of an mRNA encoding human hsp47 (e.g., SEQ ID NO: 1).

In certain embodiments, the sequence of the antisense strand that iscomplementary to a sequence of an mRNA encoding human hsp47 includes asequence complimentary to a sequence between nucleotides 600-800; or801-899; or 900-1000; or 1001-1300 of SEQ ID NO: 1; or betweennucleotides 650-730; or 900-975 of SEQ ID NO: 1. In some embodiments,the antisense strand includes a sequence that is complementary to asequence of an mRNA encoding human hsp47 corresponding to nucleotides674-693 of SEQ ID NO: 1 or a portion thereof; or nucleotides 698-716 ofSEQ ID NO: 1 or a portion thereof; or nucleotides 698-722 of SEQ ID NO:1 or a portion thereof; or nucleotides 701-720 of SEQ ID NO: 1 or aportion thereof; or nucleotides 920-939 of SEQ ID NO: 1 or a portionthereof; or nucleotides 963-982 of SEQ ID NO: 1 or a portion thereof; ornucleotides 947-972 of SEQ ID NO: 1 or a portion thereof; or nucleotides948-966 of SEQ ID NO: 1 or a portion thereof; or nucleotides 945-969 ofSEQ ID NO: 1 or a portion thereof; or nucleotides 945-963 of SEQ ID NO:1 or a portion thereof.

In certain embodiments, the antisense strand of a nucleic acid molecule(e.g., a siNA molecule) as disclosed herein includes a sequencecorresponding to SEQ ID NO: 4 or a portion thereof; or SEQ ID NO: 6 or aportion thereof; or SEQ ID NO: 8 or a portion thereof; or SEQ ID NO: 10or a portion thereof; or SEQ ID NO: 12 or a portion thereof; or SEQ IDNO: 14 or a portion thereof; or SEQ ID NO: 16 or a portion thereof; orSEQ ID NO: 18 or a portion thereof; or SEQ ID NO: 20 or a portionthereof; or SEQ ID NO: 22 or a portion thereof; or SEQ ID NO: 24 or aportion thereof; or SEQ ID NO: 26 or a portion thereof; or SEQ ID NO: 28or a portion thereof; or SEQ ID NO: 30 or a portion thereof; or SEQ IDNO: 32 or a portion thereof; or SEQ ID NO: 34 or a portion thereof; orSEQ ID NO: 36 or a portion thereof; or SEQ ID NO: 38 or a portionthereof; or SEQ ID NO: 40 or a portion thereof; or SEQ ID NO: 42 or aportion thereof; or SEQ ID NO: 44 or a portion thereof; or SEQ ID NO: 46or a portion thereof; or SEQ ID NO: 48 or a portion thereof; or SEQ IDNO: 50 or a portion thereof; or SEQ ID NO: 52 or a portion thereof; orSEQ ID NO: 54 or a portion thereof; or SEQ ID NO: 56 or a portionthereof; or SEQ ID NO: 58 or a portion thereof. In certain embodiments,the sense strand of a nucleic acid molecule (e.g., a siNA molecule) asdisclosed herein includes a sequence corresponding to SEQ ID NO: 3 or aportion thereof; or SEQ ID NO: 5 or a portion thereof; or SEQ ID NO: 7or a portion thereof; or SEQ ID NO: 9 or a portion thereof; or SEQ IDNO: 11 or a portion thereof; or SEQ ID NO: 13 or a portion thereof; orSEQ ID NO: 15 or a portion thereof; or SEQ ID NO: 17 or a portionthereof; or SEQ ID NO: 19 or a portion thereof; or SEQ ID NO: 21 or aportion thereof; or SEQ ID NO: 23 or a portion thereof; or SEQ ID NO: 25or a portion thereof; or SEQ ID NO: 27 or a portion thereof; or SEQ IDNO: 29 or a portion thereof; or SEQ ID NO: 31 or a portion thereof; orSEQ ID NO: 33 or a portion thereof; or SEQ ID NO: 35 or a portionthereof; or SEQ ID NO: 37 or a portion thereof; or SEQ ID NO: 39 or aportion thereof; or SEQ ID NO: 41 or a portion thereof; or SEQ ID NO: 43or a portion thereof; or SEQ ID NO: 45 or a portion thereof; or SEQ IDNO: 47 or a portion thereof; or SEQ ID NO: 49 or a portion thereof; orSEQ ID NO: 51 or a portion thereof; or SEQ ID NO: 53 or a portionthereof; or SEQ ID NO: 55 or a portion thereof; or SEQ ID NO: 57 or aportion thereof.

In certain preferred embodiments, the antisense strand of a nucleic acidmolecule (e.g., a siNA molecule) as disclosed herein includes a sequencecorresponding to any one of the antisense sequences shown on Table A-19.In certain preferred embodiments the antisense strand and the strand areselected from the sequence pairs shown in Table A-19. In someembodiments the antisense and sense strands are selected from thesequence pairs set forth in SERPINH1_(—)4, SERPINH1_(—)12,SERPINH1_(—)18, SERPINH1_(—)30, SERPINH1_(—)58 and SERPINH1_(—)88. Insome embodiments the antisense and sense strands are selected from thesequence pairs set forth in SERPINH1_(—)4 (SEQ ID NOS:195 and 220),SERPINH1_(—)12 (SEQ ID NOS:196 and 221), SERPINH1_(—)30 (SEQ ID NOS:199and 224), and SERPINH1_(—)58 (SEQ ID NOS:208 and 233).

In some embodiments the antisense and sense strands of a nucleic acidmolecule (e.g., a siNA molecule) as disclosed herein includes thesequence pairs set forth in SERPINH1_(—)4 (SEQ ID NOS:195 and 220). Insome embodiments of a nucleic acid molecule (e.g., a siNA molecule) asdisclosed herein includes the antisense and sense strands of thesequence pairs set forth in SERPINH1_(—)12 (SEQ ID NOS:196 and 221). Insome embodiments the antisense and sense strands of a nucleic acidmolecule (e.g., a siNA molecule) as disclosed herein includes thesequence pairs set forth in SERPINH1_(—)30 (SEQ ID NOS:199 and 224). Insome embodiments of a nucleic acid molecule (e.g., a siNA molecule) asdisclosed herein includes the antisense and sense strands of thesequence pairs set forth in SERPINH1_(—)58 (SEQ ID NOS:208 and 233).

In certain embodiments, the antisense strand of a nucleic acid molecule(e.g., a siNA molecule) as disclosed herein includes a sequencecorresponding to any one of the antisense sequences shown on any one ofTables B or C.

In certain preferred embodiments, the antisense strand of a nucleic acidmolecule (e.g., a siNA molecule) as disclosed herein includes a sequencecorresponding to any one of the antisense sequences shown on Table A-18.In certain preferred embodiments the antisense strand and the strand areselected from the sequence pairs shown in Table A-18. In someembodiments of a nucleic acid molecule (e.g., a siNA molecule) asdisclosed herein includes the antisense and sense strands selected fromthe sequence pairs set forth in SERPINH1_(—)2 (SEQ ID NOS: 60 and 127),SERPINH1_(—)6 (SEQ ID NOS: 63 and 130), SERPINH1_(—)11 (SEQ ID NOS: 68and 135), SERPINH1_(—)13 (SEQ ID NOS: 69 and 136), SERPINH1_(—)45 (SEQID NOS: 97 and 164), SERPINH1_(—)45a (SEQ ID NOS: 98 and 165),SERPINH1_(—)51 (SEQ ID NOS: 101 and 168), SERPINH1_(—)52 (SEQ ID NOS:102and 169) or SERPINH1_(—)86 (SEQ ID NOS: 123 and 190). In some preferredembodiments the antisense and sense strands are selected from thesequence pairs set forth in SERPINH1_(—)2 (SEQ ID NOS: 60 and 127),SERPINH1_(—)6 (SEQ ID NOS: 63 and 130). SERPINH1_(—)45a (SEQ ID NOS: 98and 165), and SERPINH1_(—)51 (SEQ ID NOS: 101 and 168).

In some preferred embodiments of a nucleic acid molecule (e.g., a siNAmolecule) as disclosed herein includes the antisense and sense strandsselected from the sequence pairs set forth in SERPINH1_(—)2 (SEQ ID NOS:60 and 127). In some embodiments the antisense and sense strands includethe sequence pairs set forth in SERPINH1_(—)6 (SEQ ID NOS: 63 and 130).In some embodiments of a nucleic acid molecule (e.g., a siNA molecule)as disclosed herein includes the antisense and sense strands of thesequence pairs set forth in SERPINH1_(—)11 (SEQ ID NOS: 68 and 135). Insome embodiments the antisense and sense strands are the sequence pairsset forth in SERPINH1_(—)13 (SEQ ID NOS: 69 and 136). In someembodiments the antisense and sense strands are the sequence pairs setforth in SERPINH1_(—)45 (SEQ ID NOS: 97 and 164). In some embodimentsthe antisense and sense strands are the sequence pairs set forth inSERPINH1_(—)45a (SEQ ID NOS: 98 and 165). In some embodiments theantisense and sense strands are the sequence pairs set forth inSERPINH1_(—)51 (SEQ ID NOS: 101 and 168).

In certain embodiments, the antisense strand of a nucleic acid molecule(e.g., a siNA molecule) as disclosed herein includes a sequencecorresponding to any one of the antisense sequences shown on any one ofTables D or E.

In various embodiments of nucleic acid molecules (e.g., siNA molecules)as disclosed herein, the antisense strand may be 15 to 49 nucleotides inlength (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,47, 48 or 49 nucleotides in length); or 17-35 nucleotides in length; or17-30 nucleotides in length; or 15-25 nucleotides in length; or 18-25nucleotides in length; or 18-23 nucleotides in length; or 19-21nucleotides in length; or 25-30 nucleotides in length; or 26-28nucleotides in length. In some embodiments of nucleic acid molecules(e.g., siNA molecules) as disclosed herein, the antisense strand may be19 nucleotides in length Similarly the sense strand of nucleic acidmolecules (e.g., siNA molecules) as disclosed herein may be 15 to 49nucleotides in length (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48 or 49 nucleotides in length); or 17-35 nucleotides inlength; or 17-30 nucleotides in length; or 15-25 nucleotides in length;or 18-25 nucleotides in length; or 18-23 nucleotides in length; or 19-21nucleotides in length; or 25-30 nucleotides in length; or 26-28nucleotides in length. In some embodiments of nucleic acid molecules(e.g., siNA molecules) as disclosed herein, the sense strand may be 19nucleotides in length. In some embodiments of nucleic acid molecules(e.g., siNA molecules) as disclosed herein, the antisense strand and thesense strand may be 19 nucleotides in length. The duplex region of thenucleic acid molecules (e.g., siNA molecules) as disclosed herein may be15-49 nucleotides in length (e.g., about 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48 or 49 nucleotides in length), 15-35nucleotides in length; or 15-30 nucleotides in length; or about 15-25nucleotides in length; or 17-25 nucleotides in length; or 17-23nucleotides in length; or 17-21 nucleotides in length; or 25-30nucleotides in length; or 25-28 nucleotides in length. In variousembodiments of nucleic acid molecules (e.g., siNA molecules) asdisclosed herein, the duplex region may be 19 nucleotides in length.

In certain embodiments, the sense and antisense strands of a nucleicacid (e.g., an siNA nucleic acid molecule) as provided herein areseparate polynucleotide strands. In some embodiments, the separateantisense and sense strands form a double stranded structure viahydrogen bonding, for example, Watson-Crick base pairing. In someembodiments the sense and antisense strands are two separate strandsthat are covalently linked to each other. In other embodiments, thesense and antisense strands are part of a single polynucleotide strandhaving both a sense and antisense region; in some preferred embodimentsthe polynucleotide strand has a hairpin structure.

In certain embodiments, the nucleic acid molecule (e.g., siNA molecule)is a double stranded nucleic acid (dsNA) molecule that is symmetricalwith regard to overhangs, and has a blunt end on both ends. In otherembodiments the nucleic acid molecule (e.g., siNA molecule) is a dsNAmolecule that is symmetrical with regard to overhangs, and has anoverhang on both ends of the dsNA molecule; preferably the molecule hasoverhangs of 1, 2, 3, 4, 5, 6, 7, or 8 nucleotides; preferably themolecule has 2 nucleotide overhangs. In some embodiments the overhangsare 5′ overhangs; in alternative embodiments the overhangs are 3′overhangs. In certain embodiments, the overhang nucleotides are modifiedwith modifications as disclosed herein. In some embodiments the overhangnucleotides are 2′-deoxynucleotides.

In certain preferred embodiments, the nucleic acid molecule (e.g., siNAmolecule) is a dsNA molecule that is asymmetrical with regard tooverhangs, and has a blunt end on one end of the molecule and anoverhang on the other end of the molecule. In certain embodiments theoverhang is 1, 2, 3, 4, 5, 6, 7, or 8 nucleotides; preferably theoverhang is 2 nucleotides. In some preferred embodiments an asymmetricaldsNA molecule has a 3′-overhang (for example a two nucleotide3′-overhang) on one side of a duplex occurring on the sense strand; anda blunt end on the other side of the molecule. In some preferredembodiments an asymmetrical dsNA molecule has a 5′-overhang (for examplea two nucleotide 5′-overhang) on one side of a duplex occurring on thesense strand; and a blunt end on the other side of the molecule. Inother preferred embodiments an asymmetrical dsNA molecule has a3′-overhang (for example a two nucleotide 3′-overhang) on one side of aduplex occurring on the antisense strand; and a blunt end on the otherside of the molecule. In some preferred embodiments an asymmetrical dsNAmolecule has a 5′-overhang (for example a two nucleotide 5′-overhang) onone side of a duplex occurring on the antisense strand; and a blunt endon the other side of the molecule. In certain preferred embodiments, theoverhangs are 2′-deoxynucleotides.

In some embodiments, the nucleic acid molecule (e.g., siNA molecule) hasa hairpin structure (having the sense strand and antisense strand on onepolynucleotide), with a loop structure on one end and a blunt end on theother end. In some embodiments, the nucleic acid molecule has a hairpinstructure, with a loop structure on one end and an overhang end on theother end (for example a 1, 2, 3, 4, 5, 6, 7, or 8 nucleotide overhang);in certain embodiments, the overhang is a 3′-overhang; in certainembodiments the overhang is a 5′-overhang; in certain embodiments theoverhang is on the sense strand; in certain embodiments the overhang ison the antisense strand.

In some preferred embodiments, the nucleic acid molecule is selectedfrom the nucleic acid molecules shown on Table I.

The nucleic acid molecules (e.g., siNA molecule) disclosed herein mayinclude one or more modifications or modified nucleotides such asdescribed herein. For example, a nucleic acid molecule (e.g., siNAmolecule) as provided herein may include a modified nucleotide having amodified sugar; a modified nucleotide having a modified nucleobase; or amodified nucleotide having a modified phosphate group. Similarly, anucleic acid molecule (e.g., siNA molecule) as provided herein mayinclude a modified phosphodiester backbone and/or may include a modifiedterminal phosphate group.

Nucleic acid molecules (e.g., siNA molecules) as provided may have oneor more nucleotides that include a modified sugar moiety, for example asdescribed herein. In some preferred embodiments the modified sugarmoiety is selected from the group consisting of 2′-O-methyl,2′-methoxyethoxy, 2′-deoxy, 2′-fluoro, 2′-allyl,2′-O-[2-(methylamino)-2-oxoethyl], 4′-thio, 4′-(CH₂)₂—O-2′-bridge,2′-locked nucleic acid, and 2′-O—(N-methyl carbamate).

Nucleic acid molecules (e.g., siNA molecules) as provided may have oneor more modified nucleobase(s) for example as described herein, whichpreferably may be one selected from the group consisting of xanthine,hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives ofadenine and guanine, 2-propyl and other alkyl derivatives of adenine andguanine, 5-halouracil and cytosine, 5-propynyl uracil and cytosine,6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil),4-thiouracil, 8-halo, amino, thiol, thioalkyl, hydroxyl and other8-substituted adenines and guanines, 5-trifluoromethyl and other5-substituted uracils and cytosines, 7-methyl guanine, andacyclonucleotides.

Nucleic acid molecules (e.g., siNA molecules) as provided may have oneor more modifications to the phosphodiester backbone, for example asdescribed herein. In some preferred embodiments the phosphodiester bondis modified by substituting the phosphodiester bond with aphosphorothioate, 3′-(or -5′)deoxy-3′-(or -5′)thio-phosphorothioate,phosphorodithioate, phosphoroselenates, 3′-(or -5′)deoxy phosphinates,borano phosphates, 3′-(or -5′)deoxy-3′-(or 5′-)amino phosphoramidates,hydrogen phosphonates, borano phosphate esters, phosphoramidates, alkylor aryl phosphonates and phosphotriester or phosphorus linkages.

In various embodiments, the provided nucleic acid molecules (e.g., siNAmolecules) may include one or modifications in the sense strand but notthe antisense strand. In some embodiments the provided nucleic acidmolecules (e.g., siNA molecules) include one or more modifications inthe antisense strand but not the sense strand. In some embodiments theprovided nucleic acid molecules (e.g., siNA molecules) include one ormore modifications in the both the sense strand and the antisensestrand.

In some embodiments in which the provided nucleic acid molecules (e.g.,siNA molecules) have modifications, the sense strand includes a patternof alternating modified and unmodified nucleotides, and/or the antisensestrand includes a pattern of alternating modified and unmodifiednucleotides; in some preferred versions of such embodiments themodification is a 2′-O-methyl (2′ methoxy or 2′OMe) sugar moiety. Thepattern of alternating modified and unmodified nucleotides may startwith a modified nucleotide at the 5′ end or 3′ end of one of thestrands; for example the pattern of alternating modified and unmodifiednucleotides may start with a modified nucleotide at the 5′ end or 3′ endof the sense strand and/or the pattern of alternating modified andunmodified nucleotides may start with a modified nucleotide at the 5′end or 3′ end of the antisense strand. When both the antisense and sensestrand include a pattern of alternating modified nucleotides, thepattern of modified nucleotides may be configured such that modifiednucleotides in the sense strand are opposite modified nucleotides in theantisense strand; or there may be a phase shift in the pattern such thatmodified nucleotides of the sense strand are opposite unmodifiednucleotides in the antisense strand and vice-versa.

The nucleic acid molecules (e.g., siNA molecules) as provided herein mayinclude 1-3 (i.e., 1, 2 or 3) deoxynucleotides at the 3′ end of thesense and/or antisense strand.

The nucleic acid molecules (e.g., siNA molecules) as provided herein mayinclude a phosphate group at the 5′ end of the sense and/or antisensestrand.

In one aspect, provided are double stranded nucleic acid moleculeshaving the structure (A1):

(A1) 5′ (N)x-Z 3′ (antisense strand)

-   -   3′ Z′—(N′)y-z″ 5′ (sense strand)        wherein each of N and N′ is a nucleotide which may be unmodified        or modified, or an unconventional moiety;        wherein each of (N)x and (N′)y is an oligonucleotide in which        each consecutive N or N′ is joined to the next N or N′ by a        covalent bond;        wherein each of Z and Z′ is independently present or absent, but        if present independently includes 1-5 consecutive nucleotides or        non-nucleotide moieties or a combination thereof covalently        attached at the 3′ terminus of the strand in which it is        present;        wherein z″ may be present or absent, but if present is a capping        moiety covalently attached at the 5′ terminus of (N′)y;        wherein each of x and y is independently an integer between 18        and 40;        wherein the sequence of (N′)y has complementary to the sequence        of (N)x; and wherein (N)x includes an antisense sequence to SEQ        ID NO:1. In some embodiments (N)x includes an antisense        oligonucleotide present in Table A-19. In other embodiments (N)x        is selected from an antisense oligonucleotide present in Tables        B or C.

In some embodiments the covalent bond joining each consecutive N or N′is a phosphodiester bond.

In some embodiments x=y and each of x and y is 19, 20, 21, 22 or 23. Invarious embodiments x=y=19.

In some embodiments of nucleic acid molecules (e.g., siNA molecules) asdisclosed herein, the double stranded nucleic acid molecule is a siRNA,siNA or a miRNA.

In some embodiments, the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)4 (SEQ ID NOS:195 and 220),SERPINH1_(—)12 (SEQ ID NOS:196 and 221), SERPINH1_(—)30 (SEQ ID NOS:199and 224), and SERPINH1_(—)58 (SEQ ID NOS:208 and 233),

In some embodiments the antisense and sense strands are the sequencepairs set forth in SERPINH1_(—)4 (SEQ ID NOS:195 and 220). In someembodiments the antisense and sense strands are the sequence pairs setforth in SERPINH1_(—)12 (SEQ ID NOS:196 and 221). In some embodimentsthe antisense and sense strands are the sequence pairs set forth inSERPINH1_(—)30 (SEQ ID NOS:199 and 224), In some embodiments theantisense and sense strands are the sequence pairs set forth inSERPINH1_(—)58 (SEQ ID NOS:208 and 233).

In some embodiments the double stranded nucleic acid molecules comprisea DNA moiety or a mismatch to the target at position 1 of the antisensestrand (5′ terminus). Such a structure is described herein. According toone embodiment provided are modified nucleic acid molecules having astructure (A2) set forth below:

(A2) 5′ N¹—(N)x-Z 3′ (antisense strand)

-   -   3′ Z′—N²—(N′)y-z″ 5′ (sense strand)        wherein each of N², N and N′ is an unmodified or modified        ribonucleotide. or an unconventional moiety;        wherein each of (N)x and (N′)y is an oligonucleotide in which        each consecutive N or N′ is joined to the adjacent N or N′ by a        covalent bond;        wherein each of x and y is independently an integer between 17        and 39;        wherein the sequence of (N′)y has complementarity to the        sequence of (N)x and (N)x has complementarity to a consecutive        sequence in a target RNA;        wherein N¹ is covalently bound to (N)x and is mismatched to the        target RNA or is a complementary DNA moiety to the target RNA;        wherein N¹ is a moiety selected from the group consisting of        natural or modified uridine, deoxyribouridine, ribothymidine,        deoxyribothymidine, adenosine or deoxyadenosine;        wherein z″ may be present or absent, but if present is a capping        moiety covalently attached at the 5′ terminus of N²—(N′)y; and        wherein each of Z and Z′ is independently present or absent, but        if present is independently 1-5 consecutive nucleotides,        consecutive non-nucleotide moieties or a combination thereof        covalently attached at the 3′ terminus of the strand in which it        is present.

In some embodiments the sequence of (N′)y is fully complementary to thesequence of (N)x. In various embodiments sequence of N²—(N′)y iscomplementary to the sequence of N¹—(N)x. In some embodiments (N)xcomprises an antisense that is fully complementary to about 17 to about39 consecutive nucleotides in a target RNA. In other embodiments (N)xcomprises an antisense that is substantially complementary to about 17to about 39 consecutive nucleotides in a target RNA.

In some embodiments N¹ and N² form a Watson-Crick base pair, In someembodiments N¹ and N² form a non-Watson-Crick base pair. In someembodiments a base pair is formed between a ribonucleotide and adeoxyribonucleotide.

In some embodiments x=y=18, x=y=19 or x=y=20. In preferred embodimentsx=y=18. When x=18 in N¹—(N)x, N¹ refers to position land positions 2-19are included in (N)₁₈. When y=18 in N²—(N′)y, N² refers to position 19and positions 1-18 are included in (N′)₁₈.

In some embodiments N¹ is covalently bound to (N)x and is mismatched tothe target RNA. In various embodiments N¹ is covalently bound to (N)xand is a DNA moiety complementary to the target RNA.

In some embodiments a uridine in position 1 of the antisense strand issubstituted with an N¹ selected from adenosine, deoxyadenosine,deoxyuridine (dU), ribothymidine or deoxythymidine. In variousembodiments N¹ selected from adenosine, deoxyadenosine or deoxyuridine.

In some embodiments guanosine in position 1 of the antisense strand issubstituted with an N¹ selected from adenosine, deoxyadenosine, uridine,deoxyuridine, ribothymidine or deoxythymidine. In various embodiments N¹is selected from adenosine, deoxyadenosine, uridine or deoxyuridine.

In some embodiments cytidine in position 1 of the antisense strand issubstituted with an N¹ selected from adenosine, deoxyadenosine, uridine,deoxyuridine, ribothymidine or deoxythymidine. In various embodiments N¹is selected from adenosine, deoxyadenosine, uridine or deoxyuridine.

In some embodiments adenosine in position 1 of the antisense strand issubstituted with an N¹ selected from deoxyadenosine, deoxyuridine,ribothymidine or deoxythymidine. In various embodiments N¹ selected fromdeoxyadenosine or deoxyuridine.

In some embodiments N¹ and N² form a base pair between uridine ordeoxyuridine, and adenosine or deoxyadenosine. In other embodiments N¹and N² form a base pair between deoxyuridine and adenosine.

In some embodiments the double stranded nucleic acid molecule is asiRNA, siNA or a miRNA. The double stranded nucleic acid molecules asprovided herein are also referred to as “duplexes”.

In some embodiments (N)x includes and antisense oligonucleotide presentin Table A-18. In some embodiments x=y=18 and N1-(N)x includes anantisense oligonucleotide present in Table A-18. In some embodimentsx=y=19 or x=y=20. In certain preferred embodiments x=y=−18. In someembodiments x=y=18 and the sequences of N1-(N)x and N2-(N′)y areselected from the pair of oligonucleotides set forth in Table A-18. Insome embodiments x=y=18 and the sequences of N1-(N)x and N2-(N′)y areselected from the pair of oligonucleotides set forth in Tables D and E.In some embodiments the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)2 (SEQ ID NOS: 60 and 127),SERPINH1_(—)6 (SEQ ID NOS: 63 and 130), SERPINH1_(—)11 (SEQ ID NOS: 68and 135), SERPINH1_(—)13 (SEQ ID NOS: 69 and 136), SERPINH1_(—)45 (SEQID NOS: 97 and 164), SERPINH1_(—)45a (SEQ ID NOS: 98 and 165),SERPINH1_(—)51 (SEQ ID NOS: 101 and 168), SERPINH1_(—)51a (SEQ ID NOS:105 and 172), SERPINH1_(—)52 (SEQ ID NOS:102 and 169) or SERPINH1_(—)86(SEQ ID NOS: 123 and 190). In some preferred embodiments the antisenseand sense strands are selected from the sequence pairs set forth inSERPINH1_(—)2 (SEQ ID NOS: 60 and 127), SERPINH1_(—)6 (SEQ ID NOS: 63and 130), SERPINH1_(—)45a (SEQ ID NOS: 98 and 165), SERPINH1_(—)51 (SEQID NOS: 101 and 168) and SERPINH1_(—)51a (SEQ ID NOS: 105 and 172).

In some preferred embodiments the antisense and sense strands areselected from the sequence pairs set forth in SERPINH1_(—)2 (SEQ ID NOS:60 and 127). In some embodiments the antisense and sense strands are thesequence pairs set forth in SERPINH1_(—)6 (SEQ ID NOS: 63 and 130). Insome embodiments the antisense and sense strands are the sequence pairsset forth in SERPINH1_(—)11(SEQ ID NOS: 68 and 135). In some embodimentsthe antisense and sense strands are the sequence pairs set forth inSERPINH1_(—)13 (SEQ ID NOS: 69 and 136). In some embodiments theantisense and sense strands are the sequence pairs set forth inSERPINH1_(—)45 (SEQ ID NOS: 97 and 164). In some embodiments theantisense and sense strands are the sequence pairs set forth inSERPINH1_(—)45a (SEQ ID NOS: 98 and 165). In some embodiments theantisense and sense strands are the sequence pairs set forth inSERPINH1_(—)51 (SEQ ID NOS: 101 and 168). In some embodiments theantisense and sense strands are the sequence pairs set forth inSERPINH1_(—)51a (SEQ ID NOS: 105 and 172). In some embodiments theantisense and sense strands are the sequence pairs set forth inSERPINH1_(—)52 (SEQ ID NOS:102 and 169). In some embodiments theantisense and sense strands are the sequence pairs set forth in (SEQ IDNOS: 123 and 190). In some preferred embodiments the antisense and sensestrands are selected from the sequence pairs set forth in SERPINH1_(—)2(SEQ ID NOS: 60 and 127), SERPINH1_(—)6 (SEQ ID NOS: 63 and 130),SERPINH1_(—)45a (SEQ ID NOS: 98 and 165), SERPINH1_(—)51 (SEQ ID NOS:101 and 168) and SERPINH1_(—)51a (SEQ ID NOS: 105 and 172).

In some embodiments N1 and N2 form a Watson-Crick base pair. In otherembodiments N1 and N2 form a non-Watson-Crick base pair. In someembodiments N1 is a modified riboadenosine or a modified ribouridine.

In some embodiments N1 and N2 form a Watson-Crick base pair. In otherembodiments N1 and N2 form a non-Watson-Crick base pair. In certainembodiments N1 is selected from the group consisting of riboadenosine,modified riboadenosine, deoxyriboadenosine, modified deoxyriboadenosine.In other embodiments N1 is selected from the group consisting ofribouridine, deoxyribouridine, modified ribouridine, and modifieddeoxyribouridine,

In certain embodiments position 1 in the antisense strand (5′ terminus)includes deoxyribouridine (dU) or adenosine. In some embodiments N1 isselected from the group consisting of riboadenosine, modifiedriboadenosine, deoxyriboadenosine, modified deoxyriboadenosine and N2 isselected from the group consisting of ribouridine, deoxyribouridine,modified ribouridine, and modified deoxyribouridine. In certainembodiments N1 is selected from the group consisting of riboadenosineand modified riboadenosine and N2 is selected from the group consistingof ribouridine and modified ribouridine.

In certain embodiments N1 is selected from the group consisting ofribouridine, deoxyribouridine, modified ribouridine, and modifieddeoxyribouridine and N2 is selected from the group consisting ofriboadenosine, modified riboadenosine, deoxyriboadenosine, modifieddeoxyriboadenosine. In certain embodiments N1 is selected from the groupconsisting of ribouridine and deoxyribouridine and N2 is selected fromthe group consisting of riboadenosine and modified riboadenosine. Incertain embodiments N1 is ribouridine and N2 is riboadenosine. Incertain embodiments N1 is deoxyribouridine and N2 is riboadenosine.

In some embodiments of Structure (A2), N1 includes 2′OMe sugar-modifiedribouracil or 2′OMe sugar-modified riboadenosine. In certain embodimentsof structure (A), N2 includes a 2′OMe sugar modified ribonucleotide ordeoxyribonucleotide.

In some embodiments of Structure (A2), N1 includes 2′OMe sugar-modifiedribouracil or 2′OMe sugar-modified ribocytosine. In certain embodimentsof structure (A), N2 includes a 2′OMe sugar modified ribonucleotide.

In some embodiments each of N and N′ is an unmodified nucleotide. Insome embodiments at least one of N or N′ includes a chemically modifiednucleotide or an unconventional moiety. In some embodiments theunconventional moiety is selected from a mirror nucleotide, an abasicribose moiety and an abasic deoxyribose moiety. In some embodiments theunconventional moiety is a mirror nucleotide, preferably an L-DNAmoiety. In some embodiments at least one of N or N′ includes a 2′OMesugar-modified ribonucleotide.

In some embodiments the sequence of (N′)y is fully complementary to thesequence of (N)x. In other embodiments the sequence of (N′)y issubstantially complementary to the sequence of (N)x.

In some embodiments (N)x includes an antisense sequence that is fullycomplementary to about 17 to about 39 consecutive nucleotides in atarget mRNA. In other embodiments (N)x includes an antisense that issubstantially complementary to about 17 to about 39 consecutivenucleotides in a target mRNA.

In some embodiments of Structure A1 and Structure A2 the compound isblunt ended, for example wherein both Z and Z′ are absent. In analternative embodiment, at least one of Z or Z′ is present. Z and Z′independently include one or more covalently linked modified and orunmodified nucleotides, including deoxyribonucleotides andribonucleotides, or an unconventional moiety for example inverted abasicdeoxyribose moiety or abasic ribose moiety; a non-nucleotide C3, C4 orC5 moiety, an amino-6 moiety, a mirror nucleotide and the like. In someembodiments each of Z and Z′ independently includes a C3 moiety or anamino-C6 moiety. In some embodiments Z′ is absent and Z is present andincludes a non-nucleotide C3 moiety. In some embodiments Z is absent andZ′ is present and includes a non-nucleotide C3 moiety.

In some embodiments of Structure A1 and Structure A2, each N consists ofan unmodified ribonucleotide. In some embodiments of Structure A1 andStructure A2, each N′ consists of an unmodified nucleotide. In preferredembodiments, at least one of N and N′ is a modified ribonucleotide or anunconventional moiety.

In other embodiments the compound of Structure A1 or Structure A2includes at least one ribonucleotide modified in the sugar residue. Insome embodiments the compound includes a modification at the 2′ positionof the sugar residue. In some embodiments the modification in the 2′position includes the presence of an amino, a fluoro, an alkoxy or analkyl moiety. In certain embodiments the 2′ modification includes analkoxy moiety. In preferred embodiments the alkoxy moiety is a methoxymoiety (also known as 2′-O-methyl; 2′OMe; 2′-OCH3). In some embodimentsthe nucleic acid compound includes 2′OMe sugar modified alternatingribonucleotides in one or both of the antisense and the sense strands.In other embodiments the compound includes 2′OMe sugar modifiedribonucleotides in the antisense strand, (N)x or N1-(N)x, only. Incertain embodiments the middle ribonucleotide of the antisense strand;e.g. ribonucleotide in position 10 in a 19-mer strand is unmodified. Invarious embodiments the nucleic acid compound includes at least 5alternating 2′OMe sugar modified and unmodified ribonucleotides. Inadditional embodiments the compound of Structure A1 or Structure A2includes modified ribonucleotides in alternating positions wherein eachribonucleotide at the 5′ and 3′ termini of (N)x or N1-(N)x are modifiedin their sugar residues, and each ribonucleotide at the 5′ and 3′termini of (N′)y or N2-(N)y are unmodified in their sugar residues.

In some embodiments the double stranded molecule includes one or more ofthe following modifications

a) N in at least one of positions 5, 6, 7, 8, or 9 from the 5′ terminusof the antisense strand is selected from a 2′5′ nucleotide or a mirrornucleotide;b) N′ in at least one of positions 9 or 10 from the 5′ terminus of thesense strand is selected from a 2′5′ nucleotide and a pseudoUridine; andc) N′ in 4, 5, or 6 consecutive positions at the 3′ terminus positionsof (N′)y comprises a 2′5′ nucleotide.

In some embodiments the double stranded molecule includes a combinationof the following modifications

a) the antisense strand includes a 2′5′ nucleotide or a mirrornucleotide in at least one of positions 5, 6, 7, 8, or 9 from the 5′terminus; andb) the sense strand includes at least one of a 2′5′ nucleotide and apseudoUridine in positions 9 or 10 from the 5′ terminus.

In some embodiments the double stranded molecule includes a combinationof the following modifications

a) the antisense strand includes a 2′5′ nucleotide or a mirrornucleotide in at least one of positions 5, 6, 7, 8, or 9 from the 5′terminus; andc) the sense strand includes 4, 5, or 6 consecutive 2′5′ nucleotides atthe 3′ penultimate or 3′ terminal positions.

In some embodiments, the sense strand [(N)x or N1-(N)x] includes 1, 2,3, 4, 5, 6, 7, 8, or 9 2′OMe sugar modified ribonucleotides. In someembodiments, the antisense strand includes 2′OMe modifiedribonucleotides at positions 2, 4, 6, 8, 11, 13, 15, 17 and 19. In otherembodiments antisense strand includes 2′OMe modified ribonucleotides atpositions 1, 3, 5, 7, 9, 11, 13, 15, 17 and 19. In other embodiments theantisense strand includes 2′OMe modified ribonucleotides at positions 3,5, 7, 9, 11, 13, 15, 17 and 19. In some embodiments the antisense strandincludes one or more 2′OMe sugar modified pyrimidines. In someembodiments all the pyrimidine nucleotides in the antisense strand are2′OMe sugar modified. In some embodiments the sense strand includes2′OMe sugar modified pyrimidines.

In some embodiments of Structure A1 and Structure A2, neither the sensestrand nor the antisense strand is phosphorylated at the 3′ and 5′termini. In other embodiments one or both of the sense strand or theantisense strand are phosphorylated at the 3′ termini.

In some embodiments of Structure A1 and Structure A2 (N)y includes atleast one unconventional moiety selected from a mirror nucleotide, a2′5′ nucleotide and a TNA. In some embodiments the unconventional moietyis a mirror nucleotide. In various embodiments the mirror nucleotide isselected from an L-ribonucleotide (L-RNA) and an L-deoxyribonucleotide(L-DNA). In preferred embodiments the mirror nucleotide is L-DNA. Incertain embodiments the sense strand comprises an unconventional moietyin position 9 or 10 (from the 5′ terminus). In preferred embodiments thesense strand includes an unconventional moiety in position 9 (from the5′ terminus). In some embodiments the sense strand is 19 nucleotides inlength and comprises 4, 5, or 6 consecutive unconventional moieties inpositions 15, (from the 5′ terminus). In some embodiments the sensestrand includes 4 consecutive 2′5′ ribonucleotides in positions 15, 16,17, and 18. In some embodiments the sense strand includes 5 consecutive2′5′ ribonucleotides in positions 15, 16, 17, 18 and 19. In variousembodiments the sense strand further comprises Z′. In some embodimentsZ′ includes a C3OH moiety or a C3Pi moiety.

In some embodiments of Structure A1 (N′)y includes at least one L-DNAmoiety. In some embodiments x=y=19 and (N′)y, consists of unmodifiedribonucleotides at positions 1-17 and 19 and one L-DNA at the 3′penultimate position (position 18). In other embodiments x=y=19 and(N′)y consists of unmodified ribonucleotides at positions 1-16 and 19and two consecutive L-DNA at the 3′ penultimate position (positions 17and 18). In various embodiments the unconventional moiety is anucleotide joined to an adjacent nucleotide by a 2′-5′ internucleotidephosphate linkage. According to various embodiments (N′)y includes 2, 3,4, 5, or 6 consecutive ribonucleotides at the 3′ terminus linked by2′-5′ internucleotide linkages. In one embodiment, four consecutivenucleotides at the 3′ terminus of (N′)y are joined by three 2′-5′phosphodiester bonds, wherein one or more of the 2′-5′ nucleotides whichform the 2′-5′ phosphodiester bonds further includes a 3′-O-methyl(3′OMe) sugar modification. Preferably the 3′ terminal nucleotide of(N′)y includes a 2′OMe sugar modification. In certain embodiments x=y=19and (N′)y includes two or more consecutive nucleotides at positions 15,16, 17, 18 and 19 include a nucleotide joined to an adjacent nucleotideby a 2′-5′ internucleotide bond (2′-5′ nucleotide). In variousembodiments the nucleotide forming the 2′-5′ internucleotide bondincludes a 3′ deoxyribose nucleotide or a 3′ methoxy nucleotide (3′ H or3′OMe in place of a 3′ OH). In some embodiments x=y=19 and (N′)yincludes 2′-5′ nucleotides at positions 15, 16 and 17 such that adjacentnucleotides are linked by a 2′-5′ internucleotide bond between positions15-16, 16-17 and 17-18; or at positions, 15, 16, 17, 18, and 19 suchthat adjacent nucleotides are linked by a 2′-5′ internucleotide bondbetween positions 15-16, 16-17, 17-18 and 18-19 and a 3′OH is availableat the 3′ terminal nucleotide or at positions 16, 17 and 18 such thatadjacent nucleotides are linked by a 2′-5′ internucleotide bond betweenpositions 16-17, 17-18 and 18-19. In some embodiments x=y=19 and (N′)yincludes 2′-5′ nucleotides at positions 16 and 17 or at positions 17 and18 or at positions 15 and 17 such that adjacent nucleotides are linkedby a 2′-5′ internucleotide bond between positions 16-17 and 17-18 orbetween positions 17-18 and 18-19 or between positions 15-16 and 17-18,respectively. In other embodiments the pyrimidine ribonucleotides (rU,rC) in (N′)y are substituted with nucleotides joined to the adjacentnucleotide by a 2′-5′ internucleotide bond. In some embodiments theantisense and sense strands are selected from the sequence pairs setforth in SERPINH1_(—)4, SERPINH1_(—)12, SERPINH1_(—)18, SERPINH1_(—)30,SERPINH1_(—)58 or SERPINH1_(—)88, and x=y=19 and (N′)y comprises fiveconsecutive nucleotides at the 3′ terminus joined by four 2′-5′linkages, specifically the linkages between the nucleotides position15-16, 16-17, 17-18 and 18-19.

In some embodiments the linkages include phosphodiester bonds. In someembodiments the antisense and sense strands are selected from thesequence pairs set forth in SERPINH1_(—)4, SERPINH1_(—)12,SERPINH1_(—)18, SERPINH1_(—)30, SERPINH1_(—)58 or SERPINH1_(—)88 andx=y=19 and (N′)y comprises five consecutive nucleotides at the 3′terminus joined by four 2′-5′ linkages and optionally further includesZ′ and z′ independently selected from an inverted abasic moiety and a C3alkyl [C3; 1,3-propanediol mono(dihydrogen phosphate)] cap. The C3 alkylcap is covalently linked to the 3′ or 5′ terminal nucleotide. In someembodiments the 3′ C3 terminal cap further comprises a 3′ phosphate. Insome embodiments the 3′ C3 terminal cap further comprises a 3′ terminalhydroxy group.

In some embodiments the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)4, SERPINH1_(—)12,SERPINH1_(—)18, SERPINH1_(—)30, SERPINH1_(—)58 or SERPINH1_(—)88 andx=y=19 and (N′)y includes an L-DNA position 18; and (N′)y optionallyfurther includes Z′ and z′ independently selected from an invertedabasic moiety and a C3 alkyl [C3; 1,3-propanediol mono(dihydrogenphosphate)] cap.

In some embodiments (N′)y includes a 3′ terminal phosphate. In someembodiments (N′)y includes a 3′ terminal hydroxyl.

In some embodiments the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)4, SERPINH1_(—)12,SERPINH1_(—)18, SERPINH1_(—)30, SERPINH1_(—)58 or SERPINH1_(—)88 andx=y=19 and (N)x includes 2′OMe sugar modified ribonucleotides atpositions 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 or at positions 2, 4, 6, 8,11, 13, 15, 17, 19. In some embodiments the antisense and sense strandsare selected from the sequence pairs set forth in SERPINH1_(—)4,SERPINH1_(—)12, SERPINH1_(—)18, SERPINH1_(—)30, SERPINH1_(—)58 andSERPINH1_(—)88 and x=y=19 and (N)x includes 2′OMe sugar modifiedpyrimidines. In some embodiments all pyrimidines in (N)x include the2′OMe sugar modification.

In some embodiments the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)2, SERPINH1_(—)6,SERPINH1_(—)11, SERPINH1_(—)13, SERPINH1_(—)45, SERPINH1_(—)45a,SERPINH1_(—)51, SERPINH51a, SERPINH1_(—)52 or SERPINH1_(—)86 and x=y=18and N2 is a riboadenosine moiety.

In some embodiments the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)2, SERPINH1_(—)6,SERPINH1_(—)11, SERPINH1_(—)13, SERPINH1_(—)45. SERPINH1_(—)45a,SERPINH1_(—)51. SERPINH51a, SERPINH1_(—)52 or SERPINH1_(—)86 and x=y=18,and N2-(N′)y includes five consecutive nucleotides at the 3′ terminusjoined by four 2′-5′ linkages, specifically the linkages between thenucleotides position 15-16, 16-17, 17-18 and 18-19. In some embodimentsthe linkages include phosphodiester bonds.

In some embodiments the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)2, SERPINH1_(—)6,SERPINH1_(—)11, SERPINH1_(—)13, SERPINH1_(—)45, SERPINH1_(—)45a ,SERPINH1_(—)51, SERPINH1_(—)51a, SERPINH1_(—)52 or SERPINH1_(—)86 andx=y=18 and N2-(N′)y includes five consecutive nucleotides at the 3′terminus joined by four 2′-5′ linkages and optionally further includesZ′ and z′ independently selected from an inverted abasic moiety and a C3alkyl [C3; 1,3-propanediol mono(dihydrogen phosphate)] cap.

In some embodiments the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)2, SERPINH1_(—)6,SERPINH1_(—)11, SERPINH1_(—)13, SERPINH1_(—)45, SERPINH1_(—)45a,SERPINH1_(—)51, SERPINH1_(—)51a, SERPINH1_(—)52 or SERPINH1_(—)86 andx=y=18 and N²—(N′)y includes an L-DNA position 18; and (N′)y optionallyfurther includes Z′ and z′ independently selected from an invertedabasic moiety and a C3 alkyl [C3; 1,3-propanediol mono(dihydrogenphosphate)] cap.

In some embodiments N²—(N′)y comprises a 3′ terminal phosphate. In someembodiments N2-(N′)y comprises a 3′ terminal hydroxyl.

In some embodiments the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)2, SERPINH1_(—)6,SERPINH1_(—)11, SERPINH1_(—)13, SERPINH1_(—)45, SERPINH1_(—)45a,SERPINH1_(—)51, SERPINH1_(—)51a, SERPINH1_(—)52 or SERPINH1_(—)86 andx=y=18 and N¹—(N)x includes 2′OMe sugar modified ribonucleotides inpositions 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 or in positions 1, 3, 5, 9,11, 13, 15, 17, 19, or in positions 3, 5, 9, 11, 13, 15, 17, or inpositions 2, 4, 6, 8, 11, 13, 15, 17, 19. In some embodiments theantisense and sense strands are selected from the sequence pairs setforth in SERPINH1_(—)2, SERPINH1_(—)6, SERPINH1_(—)11, SERPINH1_(—)13,SERPINH1_(—)45, SERPINH1_(—)45a, SERPINH1_(—)51, SERPINH1_(—)52 orSERPINH1_(—)86 and x=y=18 and N¹—(N)x includes 2′OMe sugar modifiedribonucleotides at positions 11, 13, 15, 17 and 19 (from 5′ terminus).In some embodiments the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)2, SERPINH1_(—)6,SERPINH1_(—)11, SERPINH1_(—)13, SERPINH1-45. SERPINH1_(—)45a,SERPINH1_(—)51, SERPINH1_(—)51a, SERPINH1_(—)52 or SERPINH1_(—)86 andx=y=18 and N¹—(N)x includes 2′OMe sugar modified ribonucleotides inpositions 1, 3, 5, 7, 9, 11, 13, 15, 17, 19 or in positions 3, 5, 7, 9,11, 13, 15, 17, 19. In some embodiments the antisense and sense strandsare selected from the sequence pairs set forth in SERPINH1_(—)2,SERPINH1_(—)6, SERPINH1_(—)11, SERPINH1_(—)13, SERPINH1_(—)45,SERPINH1_(—)45a , SERPINH1_(—)51, SERPINH1_(—)52 or SERPINH1_(—)86 andx=y=18 and N1-(N)x includes 2′OMe sugar modified ribonucleotides inpositions positions 2, 4, 6, 8, 11, 13, 15, 17, 19.

In some embodiments the antisense and sense strands are selected fromthe sequence pairs set forth in SERPINH1_(—)2, SERPINH1_(—)6,SERPINH1_(—)11, SERPINH1_(—)13, SERPINH1_(—)45, SERPINH1_(—)45a,SERPINH1_(—)51, SERPINH1_(—)51a, SERPINH1_(—)52 or SERPINH1_(—)86 andx=y=18 and N1-(N)x includes 2′OMe sugar modified pyrimidines. In someembodiments all pyrimidines in (N)x include the 2′OMe sugarmodification. In some embodiments the antisense strand further includesan L-DNA or a 2′-5′ nucleotide in position 5, 6 or 7 (5′>3′). In otherembodiments the antisense strand further includes a ribonucleotide whichgenerates a 2′5′ internucleotide linkage in between the ribonucleotidesin positions 5-6 or 6-7 (5′>3′)

In additional embodiments N1-(N)x further includes Z wherein Z includesa non-nucleotide overhang. In some embodiments the non-nucleotideoverhang is C3-C3 [1,3-propanediol mono(dihydrogen phosphate)]2.

In some embodiments of Structure A2, (N)y includes at least one L-DNAmoiety. In some embodiments x=y=18 and (N′)y consists of unmodifiedribonucleotides at positions 1-16 and 18 and one L-DNA at the 3′penultimate position (position 17). In other embodiments x=y=18 and(N′)y consists of unmodified ribonucleotides at position 1-15 and 18 andtwo consecutive L-DNA at the 3′ penultimate position (positions 16 and17). In various embodiments the unconventional moiety is a nucleotidejoined to an adjacent nucleotide by a 2′-5′ internucleotide phosphatelinkage. According to various embodiments (N′)y includes 2, 3, 4, 5, or6 consecutive ribonucleotides at the 3′ terminus linked by 2′-5′internucleotide linkages. In one embodiment, four consecutivenucleotides at the 3′ terminus of (N′)y are joined by three 2′-5′phosphodiester bonds, wherein one or more of the 2′-5′ nucleotides whichform the 2′-5′ phosphodiester bonds further includes a 3′-O-methyl(3′OMe) sugar modification. Preferably the 3′ terminal nucleotide of(N′)y includes a 2′OMe sugar modification. In certain embodiments x=y=18and in (N′)y two or more consecutive nucleotides at positions 14, 15,16, 17. and 18 include a nucleotide joined to an adjacent nucleotide bya 2′-5′ internucleotide bond. In various embodiments the nucleotideforming the 2′-5′ internucleotide bond includes a 3′ deoxyribosenucleotide or a 3′ methoxy nucleotide. In some embodiments x=y=18 and(N′)y includes nucleotides joined to the adjacent nucleotide by a 2′-5′internucleotide bond between positions 15-16, 16-17 and 17-18 or betweenpositions 16-17 and 17-18. In some embodiments x=y=18 and (N′)y includesnucleotides joined to the adjacent nucleotide by a 2′-5′ internucleotidebond between positions 14-15, 15-16, 16-17, and 17-18 or betweenpositions 15-16, 16-17, and 17-18 or between positions 16-17 and 17-18or between positions 17-18 or between positions 15-16 and 17-18. Inother embodiments the pyrimidine ribonucleotides (rU, rC) in (N′)y aresubstituted with nucleotides joined to the adjacent nucleotide by a2′-5′ internucleotide bond.

In some embodiments the antisense and sense strands are selected fromthe oligonucleotide pairs set forth in Table A-18 and identified hereinas SERPINH1_(—)2 (SEQ ID NOS: 60 and 127), SERPINH1_(—)6 (SEQ ID NOS: 63and 130), SERPINH1_(—)45a (SEQ ID NOS: 98 and 165), SERPINH1_(—)51 (SEQID NOS: 101 and 168) and SERPINH1_(—)51a (SEQ ID NOS: 105 and 172).

In some embodiments the double stranded nucleic acid molecule includesthe antisense strand set forth in SEQ ID NO:127 and sense strand setforth in SEQ ID NO:60; identified herein as SERPINH1_(—)2. In someembodiments the double stranded nucleic acid molecule has the structure

wherein each “|” represents base pairing between the ribonucleotides;wherein each of A, C, G, U is independently an unmodified or modifiedribonucleotide, or an unconventional moiety;wherein each of Z and Z′ is independently present or absent, but ifpresent is independently 1-5 consecutive nucleotides or non-nucleotidemoieties or a combination thereof covalently attached at the 3′ terminusof the strand in which it is present; andwherein z″ may be present or absent, but if present is a capping moietycovalently attached at the 5′ terminus of N2-(N′)y.

In some embodiments provided is a double stranded nucleic acid moleculewherein the antisense strand (SEQ ID NO:127) includes one or more 2′OMesugar modified pyrimidines and or purines, a 2′-5′ ribonucleotide inposition 5, 6, 7 or 8, and a 3′ terminal nucleotide or non-nucleotideoverhang. In some embodiments the sense strand (SEQ ID NO:60) includes 4or 5 consecutive 2′5′ nucleotides at the 3′ terminal or penultimatepositions, a nucleotide or non-nucleotide moiety covalently attached atthe 3′ terminus and a cap moiety covalently attached at the 5′ terminus.In other embodiments the sense strand (SEQ ID NO:60) includes one ormore 2′OMe primidine, a nucleotide or non-nucleotide moiety covalentlyattached at the 3′ terminus and a cap moiety covalently attached at the5′ terminus.

In some embodiments provided is a double stranded nucleic acid moleculewherein the antisense strand (SEQ ID NO:127) includes 2′OMe sugarmodified ribonucleotides at positions (5′>3′) 1, 3, 5, 9, 11, 15, 17 and19, a 2′-5′ ribonucleotide at position 7, and a C3Pi-C3OH moietycovalently attached to the 3′ terminus; and the sense strand (SEQ IDNO:60) is selected from a sense strand which includes

a) 2′-5′ ribonucleotides at positions 15, 16, 17, 18 and 19, a C3OH 3′terminal non-nucleotide overhang; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; orb) 2′-5′ ribonucleotides at positions 15, 16, 17, 18 and 19, a 3′terminal phosphate; and an inverted abasic deoxyribonucleotide moietycovalently attached at the 5′ terminus; orc) 2′OMe sugar modified ribonucleotides at positions (5′>3′) 5, 7, 13,and 16; a 2′5′ ribonucleotide at position 18; a C3-OH moiety covalentlyattached at the 3′ terminus; and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; ord) 2′OMe sugar modified ribonucleotides at positions (5′>3′) 7, 13, 16and 18; a 2′5′ ribonucleotide at position 9; a C3OH moiety covalentlyattached at the 3′ terminus; and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; ore) 2′-5′ ribonucleotides at positions 15, 16, 17, 18, and 19: a C3-Pimoiety covalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein theantisense strand (SEQ ID NO:127) includes 2′OMe sugar modifiedribonucleotides at positions (5′>3′) 1, 3, 5, 9, 11, 15, 17, 19, a 2′-5′ribonucleotide at position 7 and a C3Pi-C3OH moiety covalently attachedto the 3′ terminus; and the sense strand (SEQ ID NO:60) includes 2′-5′ribonucleotides at positions 15, 16, 17, 18, and 19: a C3 3′ terminaloverhang; and an inverted abasic deoxyribonucleotide moiety covalentlyattached at the 5′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein theantisense strand (SEQ ID NO:127) includes 2′OMe sugar modifiedribonucleotides at positions (5′>3′) 1, 3, 5, 9, 11, 15, 17, 19, a 2′-5′ribonucleotide at position 7 and a C3Pi-C3OH, 3′ terminal overhang; andthe sense strand (SEQ ID NO:60) includes 2′-5′ ribonucleotides atpositions 15, 16, 17, 18, and 19: a 3′ terminal phosphate; and aninverted abasic deoxyribonucleotide moiety covalently attached at the 5′terminus.

Provided herein is a double stranded nucleic acid molecule wherein theantisense strand (SEQ ID NO:127) includes 2′OMe sugar modifiedribonucleotides at positions (5′>3′) 1, 3, 5, 9, 11, 15, 17, 19, a 2′-5′ribonucleotide at position 7 and a C3Pi-C3OH moiety covalently attachedto the 3′ terminus; and the sense strand (SEQ ID NO:60) includes 2′OMesugar modified ribonucleotides at positions (5′>3′) 5, 7, 13, and 16; a2′-5′ ribonucleotide at position 18; a C3-OH moiety covalently attachedat the 3′ terminus; and an inverted abasic deoxyribonucleotide moietycovalently attached at the 5′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein theantisense strand (SEQ ID NO:127) includes 2′OMe sugar modifiedribonucleotides at positions (5′>3′) 1, 3, 5, 9, 11, 15, 17, 19, a 2′-5′ribonucleotide at position 7 and a C3Pi-C3OH moiety covalently attachedto the 3′ terminus; and the sense strand (SEQ ID NO:60) includes 2′OMesugar modified ribonucleotides at positions (5′>3′) 7, 13, 16 and 18; a2′-5′ ribonucleotide at position 9; a C3-OH moiety covalently attachedat the 3′ terminus; and an inverted abasic deoxyribonucleotide moietycovalently attached at the 5′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein theantisense strand (SEQ ID NO:127) includes 2′OMe sugar modifiedribonucleotides at positions (5′>3′) 1, 3, 5, 9, 11, 15, 17, 19, a 2′-5′ribonucleotide at position 7 and a C3Pi-C3OH moiety covalently attachedto the 3′ terminus; and the sense strand (SEQ ID NO:60) includes 2′-5′ribonucleotides at positions 15, 16, 17, 18, and 19: a C3-Pi moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus.

In some embodiments provided herein is a double stranded nucleic acidmolecule wherein the antisense strand (SEQ ID NO:127) includes 2′OMesugar modified ribonucleotides at positions (5′>3′) 1, 3, 5, 9, 11, 13,15, 17, 19 and a C3-C3 3′ terminal overhang; and the sense strand (SEQID NO:60) includes 2′OMc sugar modified ribonucleotides at positions(5′>3′) 7, 9, 13, 16 and 18; and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus.

In some embodiments provided herein is a double stranded nucleic acidmolecule wherein the sense strand (SEQ ID NO:60) includes 2′-5′ribonucleotides at positions 15, 16, 17, 18, and 19: a 3′ terminalphosphate and an inverted abasic deoxyribonucleotide moiety covalentlyattached at the 5′ terminus and the antisense strand (SEQ ID NO:127)includes an antisense strand selected from one of

a) 2′OMe sugar modified ribonucleotides at positions (5′>3′) 1, 3, 5, 7,9, 11, 13, 15, 17, 19 and a C3Pi-C3OH moiety covalently attached to the3′ terminus; orb) 2′OMe sugar modified ribonucleotides at positions (5′>3′) 1, 3, 6, 8,10, 12, 14, 17, 18 and a C3Pi-C3OH moiety covalently attached to the 3′terminus.

In some embodiments provided herein is a double stranded nucleic acidmolecule which includes the antisense strand set forth in SEQ ID NO:130and the sense strand set forth in SEQ ID NO:63; identified herein asSERPINH1_(—)6. In some embodiments the duplex comprises the structure

wherein each “|” represents base pairing between the ribonucleotides;wherein each of A, C, G, U is independently an unmodified or modifiedribonucleotide, or an unconventional moiety;wherein each of Z and Z′ is independently present or absent, but ifpresent is independently 1-5 consecutive nucleotides or non-nucleotidemoieties or a combination thereof covalently attached at the 3′ terminusof the strand in which it is present; andwherein z″ may be present or absent, but if present is a capping moietycovalently attached at the 5′ terminus of N2-(N′)y.

In some embodiments provided is a double stranded nucleic acid moleculewherein the sense strand (SEQ ID NO:63) includes one or more 2′OMe sugarmodified pyrimidines; a 3′ terminal nucleotide or non-nucleotideoverhang; and cap moiety covalently attached at the 5′ terminus. In someembodiments the antisense strand (SEQ ID NO:130) includes one or more2′OMe sugar modified pyrimidine, a nucleotide or non-nucleotide moietycovalently attached at the 3′ terminus and a cap moiety covalentlyattached at the 5′ terminus.

In some embodiments provided is a double stranded nucleic acid moleculewherein the sense strand (SEQ ID NO:63) includes 2′OMe sugar modifiedribonucleotides at positions (5′>3′) 2, 14 and 18; a C3OH or C3Pi moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:130) is selected from an antisensestrand which includes

a) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 3, 5, 9,11, 13, 15 and 17; a 2′-5′ ribonucleotide at position 7; and a C3Pi-C3OHmoiety covalently attached to the 3′ terminus; orb) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 3, 5, 7,9, 12, 13 and 17; a 2′-5′ ribonucleotide at position 7; and a C3Pi-C3OHmoiety covalently attached to the 3′ terminus; orc) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 3, 5, 9,11, 13, 15 and 17; a 2′-5′ ribonucleotide at position 7; and a C3Pi-C3OHmoiety covalently attached to the 3′ terminus; ord) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 3, 5, 9,11, 13, 15 and 17; a dU in position 1; a 2′-5′ ribonucleotide inposition 7; and a C3Pi-C3OH moiety covalently attached to the 3′terminus.

In some embodiments provided is a double stranded nucleic acid moleculewherein the sense strand (SEQ ID NO:63) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 2, 14 and 18; a C3-OH moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:130) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 1, 3, 5, 9, 11, 13, 15 and 17; a2′-5′ ribonucleotide in position 7; and a C3Pi-C3OH moiety covalentlyattached to the 3′ terminus.

In some embodiments provided is a duplex oligonucleotide moleculewherein the sense strand (SEQ ID NO:63) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 14 and 18 and optionally inposition 2; a C3-OH moiety covalently attached at the 3′ terminus; andan inverted abasic deoxyribonucleotide moiety covalently attached at the5′ terminus; and the antisense strand (SEQ ID NO:130) includes 2′OMesugar modified ribonucleotides in positions (5′>3′) 1, 3, 5, 7, 9, 12,13, and 17; a 2′-5′ ribonucleotide at position 7; and a C3:Pi-C3OHmoiety covalently attached to the 3′ terminus.

In some embodiments provided is a duplex oligonucleotide moleculewherein the sense strand (SEQ ID NO:63) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 14 and 18; a C3-OH moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:130) is selected from an antisensestrand which includes

a) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 3, 5, 9,11, 13, 15 and 17; a 2′-5′ ribonucleotide in position 7; and a C3Pi-C3Pior C3Pi-C3OH moiety covalently attached to the 3′ terminus; orb) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 3, 5, 7,9, 12, 13, and 17; a 2′-5′ ribonucleotide in position 7; and a C3Pi-C3Pior C3Pi-C3OH moiety covalently attached to the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:63) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 14 and 18; a C3-OH moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:130) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 1, 3, 5, 9, 11, 13, 15 and 17; a2′-5′ ribonucleotide in position 7; and a C3Pi-C3OH moiety covalentlyattached to the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:63) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 14 and 18; a C3-OH moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:130) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 1, 3, 5, 7, 9, 12, 13, and 17; a2′-5′ ribonucleotide in position 7; and a C3Pi-C3OH 3′ terminaloverhang.

In some embodiments the duplex includes the antisense strand set forthin SEQ ID NO:165 and sense strand set forth in SEQ ID NO:98; identifiedherein as SERPINH1_(—)45a. In some embodiments the duplex comprises thestructure

wherein each “|” represents base pairing between the ribonucleotides;wherein each of A, C, G, U is independently an unmodified or modifiedribonucleotide, or an unconventional moiety;wherein each of Z and Z′ is independently present or absent, but ifpresent is independently 1-5 consecutive nucleotides or non-nucleotidemoieties or a combination thereof covalently attached at the 3′ terminusof the strand in which it is present; andwherein z″ may be present or absent, but if present is a capping moietycovalently attached at the 5′ terminus of N2-(N′)y.

In some embodiments the sense strand (SEQ ID NO:98) includes 2′-5′ribonucleotides in positions (5′>3′) 15, 16, 17, and 18 or 15, 16, 17,18, and 19: a nucleotide or non-nucleotide moiety covalently attached atthe 3′ terminus, and a cap moiety covalently attached at the 5′terminus. In some embodiments the antisense strand (SEQ ID NO:165)includes 2′OMe sugar modified pyrimidine and or purines, a 2′-5′nucleotide in position 5, 6, 7, or 8 (5′>3′);, and a nucleotide ornon-nucleotide moiety covalently attached at the 3′ terminus.

In some embodiments the sense strand (SEQ ID NO:98) includes 2′-5′ribonucleotides in positions (5′>3′) 15, 16, 17, 18, and 19: a C3Pi orC3-OH 3′ terminal non-nucleotide moiety and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:165) includes an antisense strandselected from one of

a) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 2, 4, 6, 8,11, 13, 15, 17, and 19; a 2′-5′ ribonucleotide in position 7 and aC3Pi-C3Pi or C3Pi-C3OH 3′ terminal overhang; orb) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 2, 4, 6, 8,11, 13, 15, 17 and 19 and a C3Pi-C3Pi or C3Pi-C3OH 3′ terminal overhang;c) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 3, 5, 9,11, 13, 15, 17, and 19; a 2′-5′ ribonucleotide in position 7 and aC3Pi-C3Pi or C3Pi-C3OH 3′ terminal overhang; ord) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 3, 5, 7,9, 11, 13, 15, 17 and 19 and a C3Pi-C3Pi or C3Pi-C3OH 3′ terminaloverhang.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:98) includes 2′-5′ ribonucleotides in positions(5′>3′) 15, 16, 17, 18, and 19: a C3-OH 3′ terminal moiety and aninverted abasic deoxyribonucleotide moiety covalently attached at the 5′terminus; and the antisense strand (SEQ ID NO:165) includes 2′OMe sugarmodified ribonucleotides in positions (5′>3′) 2, 4, 6, 8, 11, 13, 15,17, and 19; a 2′-5′ ribonucleotide in position 7 and a C3Pi-COH 3′terminal overhang.

In some embodiments the double stranded nucleic acid molecule includesthe antisense strand set forth in SEQ ID NO:168 and sense strand setforth in SEQ ID NO:101; identified herein as SERPINH1_(—)51. In someembodiments the duplex comprises the structure

wherein each “|” represents base pairing between the ribonucleotides;wherein each of A, C, G, U is independently an unmodified or modifiedribonucleotide, or an unconventional moiety;wherein each of Z and Z′ is independently present or absent, but ifpresent is independently 1-5 consecutive nucleotides or non-nucleotidemoieties or a combination thereof covalently attached at the 3′ terminusof the strand in which it is present; andwherein z″ may be present or absent, but if present is a capping moietycovalently attached at the 5′ terminus of N2-(N′)y.

In some embodiments provided is a double stranded nucleic acid moleculewherein the sense strand (SEQ ID NO:101) includes 2′OMe sugar modifiedpyrimidines, optionally a 2′-5′ ribonucleotide in position 9 or 10; anucleotide or non-nucleotide moiety covalently attached at the 3′terminus and optionally a cap moiety covalently attached at the 5′terminus. In some embodiments the antisense strand (SEQ ID NO:168)includes 2′OMe sugar modified pyrimidine and or purines, a 2′-5′nucleotide in position 5, 6, 7, or 8 (5′>3′);, and a nucleotide ornon-nucleotide moiety covalently attached at the 3′ terminus.

In some embodiments provided is a double stranded nucleic acid moleculewherein the sense strand (SEQ ID NO:101) includes 2′OMe sugar modifiedpyrimidines in positions (5′>3′) 4, 11, 13, and 17, optionally a 2′-5′ribonucleotide in position 9 or 10, a C3Pi or C3OH non-nucleotide moietycovalently attached at the 3′ terminus and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:168) is selected from an antisensestrand which includes

a) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 8, and15, a 2′5′ ribonucleotide in position 6 or 7; a C3Pi-C3OH overhangcovalently attached at the 3′ terminus; orb) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 4, 8, 13and 15, a 2′5′ ribonucleotide in position 6 or 7; a C3Pi-C3OH overhangcovalently attached at the 3′ terminus; orc) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 4, 8, 11and 15, a 2′5′ ribonucleotide in position 6; a C3Pi-C3OH overhangcovalently attached at the 3′ terminus; ord) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 3, 8,12, 13, and 15; a 2′5′ ribonucleotide in position 6; a C3Pi-C3OH moietycovalently attached at the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:101) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 4, 11, 13, and 17, optionally a2′-5′ ribonucleotide in position 9, a C3-OH non-nucleotide moietycovalently attached at the 3′ and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; and the antisense strand(SEQ ID NO:168) includes 2′OMe sugar modified ribonucleotides inpositions (5′>3′) 1, 8, and 15, a 2′5′ ribonucleotide in position 6; aC3Pi-C3OH moiety covalently attached at the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:101) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 4, 11, 13, and 17, optionally a2′-5′ ribonucleotide in position 9, a C3-OH non-nucleotide moietycovalently attached at the 3′ and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; and the antisense strand(SEQ ID NO:168) includes 2′OMe sugar modified ribonucleotides inpositions (5′>3′) 1, 4, 8, 13 and 15, a 2′5′ ribonucleotide in position6; a C3Pi-C3OH moiety covalently attached at the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:101) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 4, 11, 13, and 17, a 2′-5′ribonucleotide in position 9, a C3OH non-nucleotide moiety covalentlyattached at the 3′ terminus and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; and the antisense strand(SEQ ID NO:168) includes 2′OMe sugar modified ribonucleotides inpositions (5′>3′) 1, 4, 8, 11 and 15, a 2′5′ ribonucleotide in position6; a C3Pi-C3OH moiety covalently attached at the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:101) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 4, 11, 13, and 17, a 2′-5′ribonucleotide in position 9, a C3OH non-nucleotide moiety covalentlyattached at the 3′ terminus and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; and the antisense strand(SEQ ID NO:168) includes 2′OMe sugar modified ribonucleotides inpositions (5′>3′) 1, 3, 8, 12, 13, and 15; a 2′5′ ribonucleotide inposition 6; a C3Pi-C3OH moiety covalently attached at the 3′ terminus.

In some embodiments the double stranded nucleic acid molecule includesthe antisense strand set forth in SEQ ID NO:168 and sense strand setforth in SEQ ID NO:101; identified herein as SERPINH1_(—)51a. In someembodiments the duplex comprises the structure

wherein each “|” represents base pairing between the ribonucleotides;wherein each of A, C, G, U is independently an unmodified or modifiedribonucleotide, or an unconventional moiety;wherein each of Z and Z′ is independently present or absent, but ifpresent is independently 1-5 consecutive nucleotides or non-nucleotidemoieties or a combination thereof covalently attached at the 3′ terminusof the strand in which it is present; and wherein z″ may be present orabsent, but if present is a capping moiety covalently attached at the 5′terminus of N2-(N′)y.

In some embodiments provided is a double stranded nucleic acid moleculewherein the sense strand (SEQ ID NO:105) includes 2′OMe sugar modifiedpyrimidines, optionally a 2′-5′ ribonucleotide in position 9 or 10; anucleotide or non-nucleotide moiety covalently attached at the 3′terminus and optionally a cap moiety covalently attached at the 5′terminus. In some embodiments the antisense strand (SEQ ID NO:172)includes 2′OMe sugar modified pyrimidine and or purines, a 2′-5′nucleotide in position 5, 6, 7, or 8 (5′>3′); and a nucleotide ornon-nucleotide moiety covalently attached at the 3′ terminus.

In some embodiments provided is a double stranded nucleic acid moleculewherein the sense strand (SEQ ID NO:105) includes 2′OMe sugar modifiedpyrimidines in positions (5′>3′) 4, 11, 13, and 17, optionally a 2′-5′ribonucleotide in position 9 or 10, a C3Pi or C3OH non-nucleotide moietycovalently attached at the 3′ terminus and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:172) is selected from an antisensestrand which includes

a) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 8, and 15,a 2′5′ ribonucleotide in position 6 or 7; a C3Pi-C3OH moiety covalentlyattached at the 3′ terminus; orb) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 4, 8, 13and 15, a 2′5′ ribonucleotide in position 6 or 7; a C3Pi-C3OH moietycovalently attached at the 3′ terminus; orc) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 4, 8, 11and 15, a 2′5′ ribonucleotide in position 6; a C3Pi-C3OH moietycovalently attached at the 3′ terminus; ord) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 3, 8, 12,13, and 15; a 2′5′ ribonucleotide in position 6; a C3Pi-C3OH moietycovalently attached at the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:105) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 4, 11, 13, and 17, optionally a2′-5′ ribonucleotide in position 9, a C3-OH non-nucleotide moietycovalently attached at the 3′ and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; and the antisense strand(SEQ ID NO:172) includes 2′OMe sugar modified ribonucleotides inpositions (5′>3′)₈, and 15, a 2′5′ ribonucleotide in position 6; aC3Pi-C3OH moiety covalently attached at the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:105) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 4, 11, 13, and 17, optionally a2′-5′ ribonucleotide in position 9, a C3-OH non-nucleotide moietycovalently attached at the 3′ and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; and the antisense strand(SEQ ID NO:172) includes 2′OMe sugar modified ribonucleotides inpositions (5′>3′) 4, 8, 13 and 15, a 2′5′ ribonucleotide in position 6;a C3Pi-C3OH moiety covalently attached at the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:105) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 4, 11, 13, and 17, a 2′-5′ribonucleotide in position 9, a C3-OH non-nucleotide moiety covalentlyattached at the 3′ terminus and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; and the antisense strand(SEQ ID NO:172) includes 2′OMe sugar modified ribonucleotides inpositions (5′>3′) 4, 8, 11 and 15, a 2′5′ ribonucleotide in position 6;a C3Pi-C3OH moiety covalently attached at the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:105) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 4, 11, 13, and 17, a 2′-5′ribonucleotide in position 9, a C3OH non-nucleotide moiety covalentlyattached at the 3′ terminus and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; and the antisense strand(SEQ ID NO:172) includes 2′OMe sugar modified ribonucleotides inpositions (5′>3″) 3, 8, 12, 13, and 15; a 2′5′ ribonucleotide inposition 6; a C3Pi-C3OH moiety covalently attached at the 3′ terminus.

In some embodiments the antisense and sense strands are selected fromthe oligonucleotide pairs set forth in Table A-19 and identified hereinas SERPINH1_(—)4 (SEQ ID NOS: 195 and 220) and SERPINH1_(—)12 (SEQ IDNOS: 196 and 221).

In some embodiments the double stranded nucleic acid molecule includesthe antisense strand set forth in SEQ ID NO:220 and sense strand setforth in SEQ ID NO:194; identified herein as SERPINH1_(—)4. In someembodiments the double stranded nucleic acid molecule has the structure

wherein each “|” represents base pairing between the ribonucleotides;wherein each of A, C, G, U is independently an unmodified or modifiedribonucleotide, or an unconventional moiety;wherein each of Z and Z′ is independently present or absent, but ifpresent is independently 1-5 consecutive nucleotides or non-nucleotidemoieties or a combination thereof covalently attached at the 3′ terminusof the strand in which it is present; andwherein z″ may be present or absent, but if present is a capping moietycovalently attached at the 5′ terminus of N2-(N′)y.

In some embodiments provided is a double stranded nucleic acid moleculewherein the antisense strand (SEQ ID NO:220) includes 2′OMe sugarmodified ribonucleotides in positions (5′>3′) 3, 5, 9, 11, 15, 17 and19, a 2′-5′ ribonucleotide in position 7, and a C3Pi-C3OH moietycovalently attached to the 3′ terminus; and the sense strand (SEQ IDNO:195) is selected from a sense strand which includes

a) 2′-5′ ribonucleotides in positions 15, 16, 17, 18 and 19, a C3OHmoiety covalently attached to the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; orb) 2′-5′ ribonucleotides in positions 15, 16, 17, 18 and 19, a 3′terminal phosphate; and an inverted abasic deoxyribonucleotide moietycovalently attached at the 5′ terminus; orc) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 5, 7, 13,and 16; a 2′5′ ribonucleotide in position 18; a C3OH moiety covalentlyattached at the 3′ terminus; and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; ord) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 7, 13, 16and 18; a 2′5′ ribonucleotide in position 9; a C3OH moiety covalentlyattached at the 3′ terminus; and an inverted abasic deoxyribonucleotidemoiety covalently attached at the 5′ terminus; ore) 2′-5′ ribonucleotides in positions 15, 16, 17, 18, and 19: a C3Pimoiety covalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein theantisense strand (SEQ ID NO:220) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 3, 5, 9, 11, 15, 17, 19, a 2′-5′ribonucleotide in position 7 and a C3Pi-C3OH moiety covalently attachedto the 3′ terminus; and the sense strand (SEQ ID NO:195) includes 2′-5′ribonucleotides in positions 15, 16, 17, 18, and 19: a C3 moietycovalently attached to the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein theantisense strand (SEQ ID NO:220) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 3, 5, 9, 11, 15, 17, 19, a 2′-5′ribonucleotide in position 7 and a C3Pi-C3OH moiety covalently attachedto the 3′ terminus; and the sense strand (SEQ ID NO:195) includes 2′-5′ribonucleotides in positions 15, 16, 17, 18, and 19: a 3′ terminalphosphate; and an inverted abasic deoxyribonucleotide moiety covalentlyattached at the 5′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein theantisense strand (SEQ ID NO:220) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 3, 5, 9, 11, 15, 17, 19, a 2′-5′ribonucleotide in position 7 and a C3Pi-C3OH moiety covalently attachedto the 3′ terminus; and the sense strand (SEQ ID NO:195) includes 2′OMesugar modified ribonucleotides in positions (5′>3′) 5, 7, 13, and 16; a2′-5′ ribonucleotide in position 18; a C3OH moiety covalently attachedat the 3′ terminus; and an inverted abasic deoxyribonucleotide moietycovalently attached at the 5′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein theantisense strand (SEQ ID NO:220) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 3, 5, 9, 11, 15, 17, 19, a 2′-5′ribonucleotide in position 7 and a C3Pi-C3OH moiety covalently attachedto the 3′ terminus; and the sense strand (SEQ ID NO:195) includes 2′OMesugar modified ribonucleotides in positions (5′>3′) 7, 13, 16 and 18; a2′-5′ ribonucleotide in position 9; a C3OH moiety covalently attached atthe 3′ terminus; and an inverted abasic deoxyribonucleotide moietycovalently attached at the 5′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein theantisense strand (SEQ ID NO:220) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 3, 5, 9, 11, 15, 17, 19, aribonucleotide in position 7 and a C3Pi-C3OH moiety covalently attachedto the 3′ terminus; and the sense strand (SEQ ID NO:195) includes 2′-5′ribonucleotides in positions 15, 16, 17, 18, and 19: a C3Pi moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus.

In some embodiments provided herein is a double stranded nucleic acidmolecule wherein the antisense strand (SEQ ID NO:220) includes 2′OMesugar modified ribonucleotides in positions (5′>3′) 1, 3, 5, 9, 11, 13,15, 17, 19 and a C3Pi-C3OH moiety covalently attached to the 3′terminus; and the sense strand (SEQ ID NO:195) includes 2′OMe sugarmodified ribonucleotides in positions (5′>3′) 7, 9, 13, 16 and 18; andan inverted abasic deoxyribonucleotide moiety covalently attached at the5′ terminus.

In some embodiments provided herein is a double stranded nucleic acidmolecule wherein the sense strand (SEQ ID NO:195) includes 2′-5′ribonucleotides in positions 15, 16, 17, 18, and 19: a 3′ terminalphosphate and an inverted abasic deoxyribonucleotide moiety covalentlyattached at the 5′ terminus and the antisense strand (SEQ ID NO:220)includes an antisense strand selected from one of

a) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 3, 5, 7, 9,11, 13, 15, 17, 19 and a C3Pi-C3OH moiety covalently attached to the 3′terminus; orb) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 3, 6, 8,10, 12, 14, 17, 18 and a C3Pi-C3OH moiety covalently attached to the 3′terminus.

In some embodiments provided herein is a double stranded nucleic acidmolecule which includes the antisense strand set forth in SEQ ID NO:130and the sense strand set forth in SEQ ID NO:63; identified herein asSERPINH1_(—)12. In some embodiments the duplex comprises the structure

wherein each “|” represents base pairing between the ribonucleotides;wherein each of A, C, G, U is independently an unmodified or modifiedribonucleotide, or an unconventional moiety;wherein each of Z and Z′ is independently present or absent, but ifpresent is independently 1-consecutive nucleotides or non-nucleotidemoieties or a combination thereof covalently attached at the 3′ terminusof the strand in which it is present; andwherein z″ may be present or absent, but if present is a capping moietycovalently attached at the 5′ terminus of N2-(N′)y.

In some embodiments provided is a double stranded nucleic acid moleculewherein the sense strand (SEQ ID NO:196) includes one or more 2′OMesugar modified pyrimidines; a 3′ terminal nucleotide or non-nucleotideoverhang; and a cap moiety covalently attached at the 5′ terminus. Insome embodiments the antisense strand (SEQ ID NO:221) includes one ormore 2′OMe sugar modified pyrimidines, a nucleotide or non-nucleotidemoiety covalently attached at the 3′ terminus, and a cap moietycovalently attached at the 5′ terminus.

In some embodiments provided is a double stranded nucleic acid moleculewherein the sense strand (SEQ ID NO:196) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 2, 14 and 18; a C3OH moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:221) is selected from an antisensestrand which includes

a) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 3, 5, 9,11, 13, 15 and 17; a 2′-5′ ribonucleotide in position 7; and a C3Pi-C3OHmoiety covalently attached to the 3′ terminus; orb) 2′OMc sugar modified ribonucleotides in positions (5′>3′) 3, 5, 7, 9,12, 13 and 17; a 2′-5′ ribonucleotide in position 7; and a C3Pi-C3OHmoiety covalently attached to the 3′ terminus.

In some embodiments provided is a double stranded nucleic acid moleculewherein the sense strand (SEQ ID NO:196) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 2, 14 and 18; a C3-OH moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:221) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 3, 5, 9, 11, 13, 15 and 17; aribonucleotide in position 7; and a C3Pi-C30H moiety covalently attachedto the 3′ terminus.

In some embodiments provided is a duplex oligonucleotide moleculewherein the sense strand (SEQ ID NO:196) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 14 and 18 and optionally inposition 2; a C3-OH moiety covalently attached at the 3′ terminus; andan inverted abasic deoxyribonucleotide moiety covalently attached at the5′ terminus; and the antisense strand (SEQ ID NO:221) includes 2′OMesugar modified ribonucleotides in positions (5′>3′) 3, 5, 7, 9, 12, 13,and 17; a 2′-5′ ribonucleotide in position 7; and a C3Pi-C3OH moietycovalently attached to the 3′ terminus.

In some embodiments provided is a duplex oligonucleotide moleculewherein the sense strand (SEQ ID NO:196) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 14 and 18; a C3-OH moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:221) is selected from an antisensestrand which includes

a) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 3, 5, 9,11, 13, 15 and 17; a 2′-5′ ribonucleotide in position 7; and a C3Pi-C3OHmoiety covalently attached to the 3′ terminus; orb) 2′OMe sugar modified ribonucleotides in positions (5′>3′) 3, 5, 7, 9,12, 13 and 17; a 2′-5′ ribonucleotide in position 7; and a C3Pi-C3OHmoiety covalently attached to the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:196) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 14 and 18; a C3-OH moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:220) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 1, 3, 5, 9, 11, 13, 15 and 17; a2′-5′ ribonucleotide in position 7; and a C3Pi-C3OH moiety covalentlyattached to the 3′ terminus.

Provided herein is a double stranded nucleic acid molecule wherein thesense strand (SEQ ID NO:196) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 14 and 18; a C3-OH moietycovalently attached at the 3′ terminus; and an inverted abasicdeoxyribonucleotide moiety covalently attached at the 5′ terminus; andthe antisense strand (SEQ ID NO:220) includes 2′OMe sugar modifiedribonucleotides in positions (5′>3′) 1, 3, 5, 7, 9, 12, 13, and 17; a2′-5′ ribonucleotide in position 7; and a C3Pi-C3OH moiety covalentlyattached to the 3′ terminus.

In further embodiments of Structures A1 and A2 (N′)y includes 1-8modified ribonucleotides wherein the modified ribonucleotide is a DNAnucleotide. In certain embodiments (N′)y includes 1, 2, 3, 4, 5, 6, 7,or up to 8 DNA moieties.

In some embodiments either Z or Z′ is present and independently includestwo non-nucleotide moieties.

In additional embodiments Z and Z′ are present and each independentlyincludes two non-nucleotide moieties.

In some embodiments each of Z and Z′ includes an abasic moiety, forexample a deoxyriboabasic moiety (referred to herein as “dAb”) orriboabasic moiety (referred to herein as “rAb”). In some embodimentseach of Z and/or Z′ includes two covalently linked abasic moieties andis for example dAb-dAb or rAb-rAb or dAb-rAb or rAb-dAb, wherein eachmoiety is covalently attached to an adjacent moiety, preferably via aphospho-based bond. In some embodiments the phospho-based bond includesa phosphorothioate, a phosphonoacetate or a phosphodiester bond. Inpreferred embodiments the phospho-based bond includes a phosphodiesterbond.

In some embodiments each of Z and/or Z′ independently includes an alkylmoiety, optionally propane [(CH2)3] moiety (C3) or a derivative thereofincluding propanol (C3-OH) and phospho derivative of propanediol(“C3-3′Pi”). In some embodiments each of Z and/or Z′ includes two alkylmoieties covalently linked to the 3′ terminus of the antisense strand orsense strand via a phosphodiester or phosphorothioate linkage andcovalently linked to one another via a phosphodiester orphosphorothioate linkage and in some examples is C3Pi-C3Pi or C3Pi-C3OH.The 3′ terminus of the antisense strand and/or the 3′ terminus of thesense strand is covalently attached to a C3 moiety via a phospho-basedbond and the C3 moiety is covalently conjugated a C3-OH moiety via aphospho-based bond. In some embodiments the phospho-based bonds includea phosphorothioate, a phosphonoacetate or a phosphodiester bond. Inpreferred embodiments the phospho-based bond includes a phosphodiesterbond.

In various embodiments of Structure A1 or Structure A2, Z and Z′ areabsent. In other embodiments Z or Z′ is present. In some embodimentseach of Z and/or Z′independently includes a C2, C3, C4, C5 or C6 alkylmoiety, optionally a C3 [propane, —(CH2)₃-] moiety or a derivativethereof including propanol (C3-OH/C3OH), propanediol, and phosphodiesterderivative of propanediol (“C3Pi”). In preferred embodiments each of Zand/or Z′ includes two hydrocarbon moieties and in some examples isC3Pi-C3OH or C3Pi-C3Pi, Each C3 is covalently conjugated to an adjacentC3 via a covalent bond, preferably a phospho-based bond. In someembodiments the phospho-based bond is a phosphorothioate, aphosphonoacetate or a phosphodiester bond.

In specific embodiments x=y=19 and Z comprises at least one C3 alkyloverhang. In some embodiments the C3-C3 overhang is covalently attachedto the 3′ terminus of (N)x or (N′)y via a covalent linkage, preferably aphosphodiester linkage. In some embodiments the linkage between a firstC3 and a second C3 is a phosphodiester linkage. In some embodiments the3′ non-nucleotide overhang is C3Pi-C3Pi. In some embodiments the 3′non-nucleotide overhang is C3Pi-C3Ps. In some embodiments the 3′non-nucleotide overhang is C3Pi-C3OH (OH is hydroxy). In someembodiments the 3′ non-nucleotide overhang is C3Pi-C3OH.

In various embodiments the alkyl moiety comprises an alkyl derivativeincluding a C3 alkyl, C4 alkyl, C5 alkyl or C6 alkyl moiety comprising aterminal hydroxyl, a terminal amino, or terminal phosphate group. Insome embodiments the alkyl moiety is a C3 alkyl or C3 alkyl derivativemoiety. In some embodiments the C3 alkyl moiety comprises propanol,propylphosphate, propylphosphorothioate or a combination thereof. The C3alkyl moiety is covalently linked to the 3′ terminus of (N′)y and/or the3′ terminus of (N)x via a phosphodiester bond. In some embodiments thealkyl moiety comprises propanol, propyl phosphate or propylphosphorothioate. In some embodiments each of Z and Z′ is independentlyselected from propanol, propyl phosphate propyl phosphorothioate,combinations thereof or multiples thereof in particular 2 or 3covalently linked propanol, propyl phosphate, propyl phosphorothioate orcombinations thereof. In some embodiments each of Z and Z′ isindependently selected from propyl phosphate, propyl phosphorothioate,propyl phospho-propanol; propyl phospho-propyl phosphorothioate;propylphospho-propyl phosphate; (propyl phosphate)₃, (propylphosphate)₂-propanol, (propyl phosphate)₂-propyl phosphorothioate. Anypropane or propanol conjugated moiety can be included in Z or Z′.

The structures of exemplary 3′ terminal non-nucleotide moieties are asfollows:

In some embodiments each of Z and Z′ is independently selected frompropanol, propyl phosphate, propyl phosphorothioate, combinationsthereof or multiples thereof.

In some embodiments each of Z and Z′ is independently selected frompropyl phosphate, propyl phosphorothioate, propyl phospho-propanol;propyl phospho-propyl phosphorothioate; propylphospho-propyl phosphate;(propyl phosphate)₃, (propyl phosphate)₂-propanol, (propylphosphate)₂-propyl phosphorothioate. Any propane or propanol conjugatedmoiety can be included in Z or Z′.

In additional embodiments each of Z and/or Z′ includes a combination ofan abasic moiety and an unmodified deoxyribonucleotide or ribonucleotideor a combination of a hydrocarbon moiety and an unmodifieddeoxyribonucleotide or ribonucleotide or a combination of an abasicmoiety (deoxyribo or ribo) and a hydrocarbon moiety. In suchembodiments, each of Z and/or Z′ includes C3-rAb or C3-dAb wherein eachmoiety is covalently bond to the adjacent moiety vi a phospho-basedbond, preferably a phosphodiester, phosphorothioate or phosphonoacetatebond.

In certain embodiments nucleic acid molecules as disclosed hereininclude a sense oligonucleotide sequence selected from any one of Oligo#s 2-67 or 68-92, shown infra in Tables A-18 and A-19, respectfully.

In certain preferred embodiments compounds provided includeCompound_(—)1, Compound_(—)2, Compound_(—)3, Compound_(—)4,Compound_(—)5, Compound_(—)6, Compound_(—)7, Compound_(—)8 andCompound_(—)9 as described herein.

In some embodiments (such as, for example, Compound_(—)1, Compound_(—)5and Compound_(—)6 as described herein) provided are 19 mer doublestranded nucleic acid molecules wherein the antisense strand is SEQ IDNO:127 and the sense strand is SEQ ID NO:60. In certain embodiments,provided are 19 mer double stranded nucleic acid molecules wherein theantisense strand is SEQ ID NO:127 and includes 2′OMe sugar modifiedribonucleotides, a ribonucleotide in at least one of positions 1, 5, 6,or 7, and a 3′ terminal non-nucleotide moiety covalently attached to the3′ terminus; and the sense strand is SEQ ID NO:60 and includes at leastone 2′5′ ribonucleotide or 2′OMe modified ribonucleotide, anon-nucleotide moiety covalently attached at the 3′ terminus and a capmoiety covalently attached at the 5′ terminus. In some embodiments,provided are 19 mer double stranded nucleic acid molecule wherein theantisense strand is SEQ ID NO:127 and includes 2′OMe sugar modifiedribonucleotides at positions 3, 5, 9, 11, 13, 15, 17, and 19 (5′>3′), a2′-5′ ribonucleotide in position 7, and a 3′ terminal C3OHnon-nucleotide moiety covalently attached at the 3′ terminus; and thesense strand is SEQ ID NO:60 and includes 5 consecutive 2′5′ribonucleotides in the 3′ terminal positions 15, 16, 17, 18, and 19(5′>3′), a C3Pi non-nucleotide moiety covalently attached at the 3′terminus and an inverted abasic moiety covalently attached at the 5′terminus.

In one embodiment provided is Compound_(—)1 that is a 19 mer doublestranded nucleic acid molecule wherein the antisense strand is SEQ IDNO:127 and includes 2′OMe sugar modified ribonucleotides at positions 3,5, 9, 11, 13, 15, 17, and 19 (5′>3′), a 2′-5′ ribonucleotide in position7, and a C3Pi-C3OH non-nucleotide moiety covalently attached at the 3′terminus; and the sense strand is SEQ ID NO:60 and includes 5consecutive 2′5′ ribonucleotides in the 3′ terminal positions 15, 16,17, 18, and 19 (5′>3′), a C3Pi non-nucleotide moiety covalently attachedat the 3′ terminus and an inverted abasic moiety covalently attached atthe 5′ terminus; and that further includes a 2′OMe sugar modifiedribonucleotide at position 1 of the antisense strand.

In one embodiment, provided is Compound_(—)6 that is a 19 mer doublestranded nucleic acid molecule wherein the antisense strand is SEQ IDNO:127 and includes 2′OMe sugar modified ribonucleotides at positions 3,5, 9, 11, 13, 15, 17, and 19 (5′>3′), a 2′-5′ ribonucleotide in position7, and a C3Pi-C3OH non-nucleotide moiety covalently attached at the 3′terminus; and the sense strand is SEQ ID NO:60 and includes 5consecutive 2′5′ ribonucleotides in the 3′ terminal positions 15, 16,17, 18, and 19 (5′>3′), a C3Pi non-nucleotide moiety covalently attachedat the 3′ terminus and an inverted abasic moiety covalently attached atthe 5′ terminus; and that further includes a 2′5′ ribonucleotide atposition 1 of the antisense strand.

In one embodiment, provided is Compound_(—)5 that is a 19 mer doublestranded nucleic acid molecule wherein the antisense strand is SEQ IDNO:127 and includes 2′OMe sugar modified ribonucleotides in positions 1,3, 5, 9, 11, 13, 15, 17, and 19 (5′>3′), a 2′-5′ ribonucleotide inposition 7, and a C3Pi-C3OH non-nucleotide moiety covalently attached atthe 3′ terminus; and the sense strand is SEQ ID NO:60 and includes 2′OMesugar modified ribonucleotides in positions (5′>3′) 7, 13, 16 and 18, a2′5′ ribonucleotide at position 9, a C3OH non-nucleotide moietycovalently attached at the 3′ terminus and an inverted abasic moietycovalently attached at the 5′ terminus.

In some embodiments (such as, for example, Compound_(—)2, andCompound_(—)7 as described herein) provided are 19 mer double strandednucleic acid molecules wherein the sense strand is SEQ ID NO:63 and theantisense strand is SEQ ID NO:130. In some embodiments provided are19-mer double stranded nucleic acid molecules wherein the sense strandis SEQ ID NO:63 and includes 2′OMe sugar modified pyrimidineribonucleotides; a non-nucleotide moiety covalently attached at the 3′terminus; and a cap moiety covalently attached at the 5′ terminus; andthe antisense strand is SEQ ID NO:130 and includes 2′OMe sugar modifiedribonucleotides; a 2′-5′ ribonucleotide at position 7; and anon-nucleotide moiety covalently attached at the 3′ terminus. In someembodiments provided are 19-mer double stranded nucleic acid moleculeswherein the sense strand is SEQ ID NO:63 and includes 2′OMe sugarmodified ribonucleotides, a non-nucleotide moiety covalently attached atthe 3′ terminus, and a cap moiety covalently attached at the 5′terminus; and the antisense strand is SEQ ID NO:130 and includes 2′OMesugar modified ribonucleotides; a ribonucleotide in at least one ofpositions 5, 6 or 7; and a non-nucleotide moiety covalently attached atthe 3′ terminus.

In one embodiment, provided is Compound_(—)2 that is a 19-mer doublestranded nucleic acid molecule wherein the sense strand is SEQ ID NO:63and includes 2′OMe sugar modified ribonucleotides in positions (5′>3′)2, 14 and 18; a C3OH moiety covalently attached at the 3′ terminus; andan inverted abasic deoxyribonucleotide moiety covalently attached at the5′ terminus; and the antisense strand is SEQ ID NO:130 and includes2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 3, 5, 9,12, 13, and 17; a 2′-5′ ribonucleotide in at least one of positions 5, 6or 7; and C3Pi-C3OH non-nucleotide moiety covalently attached at the 3′terminus.

In one embodiment, provided is Compound_(—)7 that is a 19-mer doublestranded nucleic acid molecule wherein the sense strand is SEQ ID NO:63and includes 2′OMe sugar modified ribonucleotides in positions (5′>3′)2, 14 and 18; a C3OH moiety covalently attached at the 3′ terminus; andan inverted abasic deoxyribonucleotide moiety covalently attached at the5′ terminus; and the antisense strand is SEQ ID NO:130 and includes2′OMe sugar modified ribonucleotides in positions (5′>3′) 1, 3, 5, 9,11, 13, and 17; a 2′-5′ ribonucleotide at position 7; and a C3Pi-C3OHnon-nucleotide moiety covalently attached at the 3′ terminus.

In some embodiments (such as, for example, Compound_(—)3 as describedherein) provided are 19 mer double stranded nucleic acid moleculeswherein the sense strand is SEQ ID NO:98 and the antisense strand is SEQID NO:165. In some embodiments, provided are 19-mer double strandednucleic acid molecules wherein the sense strand is SEQ ID NO:98 andincludes 2′-5′ ribonucleotides in positions at the 3′ terminus: anon-nucleotide moiety covalently attached at the 3′ terminus and a capmoiety covalently attached at the 5′ terminus; and the antisense strandis SEQ ID NO:165 and includes 2′OMe sugar modified ribonucleotides; a2′-5′ ribonucleotide in at least one of positions 5, 6 or 7 and anon-nucleotide moiety covalently attached at the 3′ terminus. In oneembodiment, provided is Compound_(—)3 that is a 19-mer double strandednucleic acid molecule wherein the sense strand is SEQ ID NO:98 andincludes 2′-5′ ribonucleotides in positions (5′>3′) 15, 16, 17, 18, and19: a C3-OH 3′ moiety covalently attached at the 3′ terminus and aninverted abasic deoxyribonucleotide moiety covalently attached at the 5′terminus; and the antisense strand is SEQ ID NO:165 and includes 2′OMesugar modified ribonucleotides in positions (5′>3′) 2, 4, 6, 8, 11, 13,15, 17, and 19; a 2′-5′ ribonucleotide in position 7 and a C3Pi-C3OHcovalently attached at the 3′ terminus.

In some embodiments (such as, for example, Compound_(—)4, Compound_(—)8and Compound_(—)9 described herein) provided are 19-mer double strandednucleic acid molecules wherein the sense strand is SEQ ID NO:101 and theantisense strand is SEQ ID NO:168. In some embodiments provided are19-mer double stranded nucleic acid molecules wherein the sense strandis SEQ ID NO:101 and includes 2′OMe sugar modified pyrimidineribonucleotides, an optional ribonucleotide in one of position 9 or 10,a non-nucleotide moiety covalently attached at the 3′ terminus and a capmoiety covalently attached at the 5′ terminus; and the antisense strandis SEQ ID NO:168 and includes 2′OMe sugar modified ribonucleotides, a2′5′ ribonucleotide in at least one of positions 5, 6, or 7; and anon-nucleotide moiety covalently attached at the 3′ terminus.

In one embodiment, provided is Compound 4 that is a 19-mer doublestranded nucleic acid molecule wherein sense strand is SEQ ID NO:101 andincludes 2′OMe sugar modified ribonucleotides in positions (5′>3′) 4,11, 13, and 17, a 2′-S′ ribonucleotide in position 9, a C3OHnon-nucleotide moiety covalently attached at the 3′ terminus and aninverted abasic deoxyribonucleotide moiety covalently attached at the 5′terminus; and the antisense strand is SEQ ID NO:168 and includes 2′OMesugar modified ribonucleotides in positions (5′>3′) 1, 4, 8, 11 and 15,a 2′5′ ribonucleotide in position 6; a 3′ C3Pi-C3OH overhang covalentlyattached at the 3′ terminus.

In one embodiment, provided is Compound_(—)8 that is a 19-mer doublestranded nucleic acid molecule wherein sense strand is SEQ ID NO:101 andincludes 2′OMe sugar modified ribonucleotides in positions (5′>3′) 4,11, 13, and 17, a C3OH non-nucleotide moiety covalently attached at the3′ terminus and an inverted abasic deoxyribonucleotide moiety covalentlyattached at the 5′ terminus; and the antisense strand is SEQ ID NO:168and includes 2′OMe sugar modified ribonucleotides in positions (5′>3′)1, 4, 8, 13 and 15. a 2′5′ ribonucleotide in position 6; and a 3′C3Pi-C3OH overhang covalently attached at the 3′ terminus.

In one embodiment, provided is Compound_(—)9 that is a 19-mer doublestranded nucleic acid molecule wherein the sense strand is SEQ ID NO:101and includes 2′OMe sugar modified ribonucleotides in positions (5′>3′)2, 4, 11, 13, and 17, a C3OH non-nucleotide moiety covalently attachedat the 3′ terminus and an inverted abasic deoxyribonucleotide moietycovalently attached at the 5′ terminus; and the antisense strand is SEQID NO:168 and includes 2′OMe sugar modified ribonucleotides in positions(5′>3′) 1, 4, 8, 11 and 15, a 2′5′ ribonucleotide in position 6; a 3′C3Pi-C3OH non-nucleotide moiety covalently attached at the 3′ terminus.

In another aspect, provided are methods for reducing the expression ofhsp47 in a cell by introducing into a cell a nucleic acid molecule asprovided herein in an amount sufficient to reduce expression of hsp47.In one embodiment, the cell is hepatocellular stellate cell. In anotherembodiment, the cell is a stellate cell in renal or pulmonary tissue. Incertain embodiments, the method is performed in vitro, in otherembodiments, the method is performed in vivo.

In yet another aspect, provided are methods for treating an individualsuffering from a disease associated with hsp47. The methods includeadministering to the individual a nucleic acid molecule such as providedherein in an amount sufficient to reduce expression of hsp47. In certainembodiments the disease associated with hsp47 is a disease selected fromthe group consisting of liver fibrosis, cirrhosis, pulmonary fibrosisincluding lung fibrosis (including ILF), any condition causing kidneyfibrosis (e.g., CKD including ESRD), peritoneal fibrosis, chronichepatic damage, fibrillogenesis, fibrotic diseases in other organs,abnormal scarring (keloids) associated with all possible types of skininjury accidental and jatrogenic (operations); scleroderma;cardiofibrosis, failure of glaucoma filtering operation; and intestinaladhesions. In some embodiments, the compounds may be useful in treatingorgan-specific indications, for example indications including thoseshown in Table 2 below:

TABLE 2 Organ Indication Skin Pathologic scarring as keloid andhypertrophic scar Surgical scarring Injury scarring keloid, ornephrogenic fibrosing dermatopathy Peritoneum Peritoneal fibrosisAdhesions Peritoneal Sclerosis associated with continual ambulatoryperitoneal dialysis (CAPD) Liver Cirrhosis including post-hepatitis Ccirrhosis, primary biliary cirrhosis Liver fibrosis, e.g. Prevention ofLiver Fibrosis in Hepatitis C carriers schistomasomiasis cholangitisLiver cirrhosis due to Hepatitis C post liver transplant orNon-Alcoholic Steatohepatitis (NASH) Pancreas inter(peri)lobularfibrosis (as in alcoholic chronic pancreatitis), periductal fibrosis (asin hereditary pancreatitis), periductal and interlobular fibrosis (as inautoimmune pancreatitis), diffuse inter- and intralobular fibrosis (asin obstructive chronic pancreatitis) Kidney Chronic Kidney Disease (CKD)of any etiology. Treatment of early stage CKD (elevated SCr) in diabeticpatients (“prevent” further deterioration in renal function) kidneyfibrosis associated with lupus glomeruloschelerosis Diabetic NephropathyHeart Congestive heart failure, Endomyocardial fibrosis, cardiofibrosisfibrosis associated with myocardial infarction Lung Asthma, Idiopathicpulmonary fibrosis (IPF); Interstitial lung fibrosis (ILF) RadiationPneumonitis leading to Pulmonary Fibrosis (e.g. due to cancer treatingradiation) Bone marrow Myeloproliferative disorders: Myelofibrosis (MF),Polycythemia vera (PV), Essential thrombocythemia (ET) idiopathicmyelofibrosis drug induced myelofibrosis. Eye Anterior segment: Cornealopacification e,g, following inherited dystrophies, herpetic keratitisor pterygia; Glaucoma Posterior segment fibrosis and traction retinaldetachment, a complication of advanced diabetic retinopathy (DR);Fibrovascular scarring and gliosis in the retina; Under the retinafibrosis for example subsequent to subretinal hemorrhage associated withneovascular AMD Retro-orbital fibrosis, postcataract surgery,proliferative vitreoretinopathy. Ocular cicatricial pemphigoid IntestineIntestinal fibrosis, Crohn's disease Vocal cord Vocal cord scarring,vocal cord mucosal fibrosis, laryngeal fibrosis VasculatureAtherosclerosis, postangioplasty arterial restenosis MultisystemicScleroderma systemic sclerosis; multifocal fibrosclerosis;sclerodermatous graft-versus-host disease in bone marrow transplantrecipients, and nephrogenic systemic fibrosis (exposure togadolinium-based contrast agents (GBCAs), 30% of MRIs) Malignancies ofMetastatic and invasive cancer by inhibiting function of activated tumorvarious origin associated myofibroblasts

In some embodiments the preferred indications include, Liver cirrhosisdue to Hepatitis C post liver transplant; Liver cirrhosis due to\Non-Alcoholic Steatohepatitis (NASH); Idiopathic Pulmonary Fibrosis;Radiation Pneumonitis leading to Pulmonary Fibrosis,; DiabeticNephropathy; Peritoneal Sclerosis associated with continual ambulatoryperitoneal dialysis (CAPD) and Ocular cicatricial pemphigoid.

Fibrotic Liver indications include Alcoholic Cirrhosis, Hepatitis Bcirrhosis, Hepatitis C cirrhosis, Hepatitis C (Hep C) cirrhosis postorthotopic liver transplant (OLTX), NASH/NAFLD, Primary biliarycirrhosis (PBC), Primary sclerosing cholangitis (PSC), Biliary atresia,alpha1 antitrypsin deficiency (A1AD), Copper storage diseases (Wilson'sdisease), Fructosemia, Galactosemia, Glycogen storage diseases(especially types III, IV, VI, IX, and X), Iron-overload syndromes(hemochromatosis), Lipid abnormalities (e.g., Gaucher's disease).Peroxisomal disorders (eg, Zellweger syndrome), Tyrosinemia, Congenitalhepatic fibrosis, Bacterial Infections (eg, brucellosis), Parasitic (eg,echinococcosis), Budd-Chiari syndrome (hepatic veno-occlusive disease).

Pulmonary Indications indications include Idiopathic Pulmonary Fibrosis,Silicosis, Pneumoconiosis, Bronchopulmonary Dysplasia in newbornfollowing neonatal respiratory distress syndrome, Bleomycin/chemo lunginjury, Brochiolitis Obliterans (BOS) post lung transplant, Chronicobstructive pulmonary disorder (COPD), Cystic Fibrosis, Asthma.

Cardiac indications include Cardiomyopathy, Atherosclerosis (Bergersdisease, etc), Endomyocardial fibrosis, Atrial Fibrillation, Scarringpost Myocardial Infarction (MI)

Other Thoracic indications include Radiation-induced capsule tissuereactions around textured breast implants and Oral submucosal fibrosis.

Renal indications include Autosomal Dominant Polycystic Kidney Disease(ADPKD), Diabetic nephropathy (diabetic glomerulosclerosis), FSGS(collapsing vs, other histologic variants), IgA Nephropathy (BergerDisease), Lupus Nephritis, Wegner's, Scleroderma, Goodpasture Syndrome,tubulointerstitial fibrosis: drug induced (protective) pencillins,cephalosporins, analgesic nephropathy, Membranoproliferativeglomerulonephritis (MPGN), Henoch-Schonlein Purpura, Congenitalnephropathies: Medullary Cystic Disease, Nail-Patella Syndrome andAlport Syndrome.

Bone Marrow indications include lympangiolyomyositosis (LAM), Chronicgraft vs. host disease, Polycythemia vera, Essential thrombocythemia,Myelofibrosis.

Ocular indications include Retinopathy of Prematurity (RoP), Ocularcicatricial pemphigoid, Lacrimal gland fibrosis, Retinal attachmentsurgery, Corneal opacity, Herpetic keratitis, Pterygia, Glaucoma,Age-related macular degeneration (AMD/ARMD), Retinal fibrosis associatedDiabetes mellitus (DM) retinopathy

Gynecological indications include Endometriosis add on to hormonaltherapy for prevention of scarring, post STD fibrosis/salphingitis,

Systemic indications include Dupuytren's disease, palmar fibromatosis,Peyronie's disease, Ledderhose disease, keloids, multifocalfibrosclerosis, nephrogenic systemic fibrosis, nephrogenic myelofibrosis(anemia).

Injury Associated Fibrotic Diseases include Bum (chemical included)induced skin & soft tissue scarring and contraction, Radiation induceskin & organ scarring post cancer therapeutic radiation treatment,Keloid (skin).

Surgical indications include peritoneal fibrosis post peritonealdialysis catheter, corneal implant, cochlear implant, other implants,silicone implants in breasts, chronic sinusitis; adhesions,pseudointimal hyperplasia of dialysis grafts.

Other indications include Chronic Pancreatitis.

In some embodiments provided is a method for treatment of a subjectsuffering from liver fibrosis comprising administering to the subject aneffective amount of a nucleic acid molecule disclosed herein, therebytreating liver fibrosis. In some embodiments the subject is sufferingfrom cirrhosis of the liver due to hepatitis. In some embodiments thesubject is suffering from cirrhosis of the liver due to NASH.

In some embodiments provided is the use of a nucleic acid moleculedisclosed herein for the manufacture of a medicament to treat liverfibrosis. In some embodiments the liver fibrosis is due to hepatitis. Insome embodiments the liver fibrosis is due to NASH.

In some embodiments provided is a method for remodeling of scar tissuecomprising administering to a subject in need thereof an effectiveamount of a nucleic acid molecule disclosed herein, thereby effectingscar tissue remodeling. In some embodiments the scar tissue is in theliver. In some embodiments the subject is suffering from cirrhosis ofthe liver due to hepatitis. In some embodiments the subject is sufferingfrom cirrhosis of the liver due to NASH.

In some embodiments provided is a method for effecting fibrosisregression comprising administering to a subject in need thereof aneffective amount of a nucleic acid molecule disclosed herein, therebyeffecting fibrosis regression.

In some embodiments provided is a method for reduction of scar tissue ina subject comprising the step of administering to the subject aneffective amount of a nucleic acid molecule disclosed herein to reducethe scar tissue. In some embodiments provided is a method for reducingscar tissue in a subject comprising the step of topically applying toscar tissue an effective amount of a nucleic acid molecule disclosedherein to reduce scar tissue.

In some embodiments provided is a method for improving the appearance ofscar tissue comprising the step of topically applying to scar tissue aneffective amount of a nucleic acid molecule disclosed herein to improvethe appearance of the scar tissue.

In some embodiments provided is a method for treatment of a subjectsuffering from lung fibrosis comprising administering to the subject aneffective amount of a nucleic acid molecule disclosed herein, therebytreating the lung fibrosis. In some embodiments the subject is sufferingfrom interstitial lung fibrosis (ILF). In some embodiments the subjectis suffering from Radiation Pneumonitis leading to Pulmonary Fibrosis.In some embodiments the subject is suffering from drug induced lungfibrosis.

In some embodiments provided is the use of a nucleic acid moleculedisclosed herein for the manufacture of a medicament to treat lungfibrosis. In some embodiments the lung fibrosis is ILF. In someembodiments the lung fibrosis drug- or radiatio-induced lung fibrosis.

In one aspect, provided are pharmaceutical compositions that include anucleic acid molecule (e.g., an siNA molecule) as described herein in apharmaceutically acceptable carrier. In certain embodiments, thepharmaceutical formulation includes, or involves, a delivery systemsuitable for delivering nucleic acid molecules (e.g., siNA molecules) toan individual such as a patient; for example delivery systems describedin more detail below.

In a related aspect, provided are compositions or kits that include anucleic acid molecule (e.g., an siNA molecule) packaged for use by apatient. The package may be labeled or include a package label or insertthat indicates the content of the package and provides certaininformation regarding how the nucleic acid molecule (e.g., an siNAmolecule) should be or can be used by a patient, for example the labelmay include dosing information and/or indications for use. In certainembodiments the contents of the label will bear a notice in a formprescribed by a government agency, for example the United States Foodand Drug administration. In certain embodiments, the label may indicatethat the nucleic acid molecule (e.g., an siNA molecule) is suitable foruse in treating a patient suffering from a disease associated withhsp47; for example, the label may indicate that the nucleic acidmolecule (e.g., an siNA molecule) is suitable for use in treatingfibroids; or for example the label may indicate that the nucleic acidmolecule (e.g., an siNA molecule) is suitable for use in treating adisease selected from the group consisting of fibrosis, liver fibrosis,cirrhosis, pulmonary fibrosis, kidney fibrosis, peritoneal fibrosis,chronic hepatic damage, and fibrillogenesis,

As used herein, the term “heat shock protein 47” or “hsp47” or “HSP47”are used interchangeably and refer to any heat shock protein 47,peptide, or polypeptide having any hsp47 protein activity. Heat shockprotein 47 is a serine proteinase inhibitor (serpin) also known, forexample, as serpin peptidase inhibitor, Glade H, member 1 (SERPINH1),SERPINH2, collagen binding protein 1 (CBP1), CBP2, gp46;arsenic-transactivated protein 3 (AsTP3); HSP47; proliferation-inducinggene 14 (PIG14); PPROM; rheumatoid arthritis antigen A-47 (RA-A47);colligin-1; and colligin-2. In certain preferred embodiments, “hsp47”refers to human hsp47. Heat shock protein 47 (or more particularly humanhsp47) may have an amino acid sequence that is the same, orsubstantially the same, as SEQ ID NO. 2 (FIG. 7).

As used herein the term “nucleotide sequence encoding hsp47” means anucleotide sequence that codes for an hsp47 protein or portion thereof.The term “nucleotide sequence encoding hsp47” is also meant to includehsp47 coding sequences such as hsp47 isoforms, mutant hsp47 genes,splice variants of hsp47 genes, and hsp47 gene polymorphisms. A nucleicacid sequence encoding hsp47 includes mRNA sequences encoding hsp47,which can also be referred to as “hsp47 mRNA.” An exemplary sequence ofhuman hsp47 mRNA is SEQ ID. NO. 1.

As used herein, the term “nucleic acid molecule” or “nucleic acid” areused interchangeably and refer to an oligonucleotide, nucleotide orpolynucleotide. Variations of “nucleic acid molecule” are described inmore detail herein. A nucleic acid molecule encompasses both modifiednucleic acid molecules and unmodified nucleic acid molecules asdescribed herein. A nucleic acid molecule may includedeoxyribonucleotides, ribonucleotides, modified nucleotides ornucleotide analogs in any combination.

As used herein, the term “nucleotide” refers to a chemical moiety havinga sugar (or an analog thereof, or a modified sugar), a nucleotide base(or an analog thereof, or a modified base), and a phosphate group (oranalog thereof, or a modified phosphate group). A nucleotide encompassesboth modified nucleotides or unmodified nucleotides as described herein.As used herein, nucleotides may include deoxyribonucleotides (e.g.,unmodified deoxyribonucleotides), ribonucleotides (e.g., unmodifiedribonucleotides), and modified nucleotide analogs including, inter alia,locked nucleic acids and unlocked nucleic acids, peptide nucleic acids,L-nucleotides (also referred to as mirror nucleotides), ethylene-bridgednucleic acid (ENA), arabinoside, PACE, nucleotides with a 6 carbonsugar, as well as nucleotide analogs (including abasic nucleotides)often considered to be non-nucleotides. In some embodiments, nucleotidesmay be modified in the sugar, nucleotide base and/or in the phosphategroup with any modification known in the art, such as modificationsdescribed herein. A “polynucleotide” or “oligonucleotide” as used hereinrefer to a chain of linked nucleotides; polynucleotides andoligonucleotides may likewise have modifications in the nucleotidesugar, nucleotide bases and phosphate backbones as are well known in theart and/or are disclosed herein.

As used herein, the term “short interfering nucleic acid”, “siNA”, or“short interfering nucleic acid molecule” refers to any nucleic acidmolecule capable of modulating gene expression or viral replication.Preferably siNA inhibits or down regulates gene expression or viralreplication. siNA includes without limitation nucleic acid moleculesthat are capable of mediating sequence specific RNAi, for example shortinterfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA),short hairpin RNA (shRNA), short interfering oligonucleotide, shortinterfering nucleic acid, short interfering modified oligonucleotide,chemically-modified siRNA, post-transcriptional gene silencing RNA(ptgsRNA), and others. As used herein, “short interfering nucleic acid”,“siNA”, or “short interfering nucleic acid molecule” has the meaningdescribed in more detail elsewhere herein.

As used herein, the term “complementary” means that a nucleic acid canform hydrogen bond(s) with another nucleic acid sequence by eithertraditional Watson-Crick or other non-traditional types. In reference tothe nucleic molecules disclosed herein, the binding free energy for anucleic acid molecule with its complementary sequence is sufficient toallow the relevant function of the nucleic acid to proceed, e.g., RNAiactivity. Determination of binding free energies for nucleic acidmolecules is well known in the art (see, e.g., Turner et al., 1987, CSHSymp. Quant. Biol. LII pp. 123-133; Frier et al., 1986, Proc. Nat. Acad.Sci. USA 83:9373-9377; Turner et al., 1987, J. Am. Chem. Soc.109:3783-3785). A percent complementarity indicates the percentage ofcontiguous residues in a nucleic acid molecule that can form hydrogenbonds (e.g., Watson-Crick base pairing) with a second nucleic acidsequence (e.g., 5, 6, 7, 8, 9, or 10 nucleotides out of a total of 10nucleotides in the first oligonucleotide being based paired to a secondnucleic acid sequence having 10 nucleotides represents 50%, 60%, 70%,80%, 90%, and 100% complementary respectively). “Fully complementary”means that all the contiguous residues of a nucleic acid sequence willhydrogen bond with the same number of contiguous residues in a secondnucleic acid sequence. In one embodiment, a nucleic acid moleculedisclosed herein includes about 15 to about 35 or more (e.g., about 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34 or 35 or more) nucleotides that are complementary to one or moretarget nucleic acid molecules or a portion thereof.

As used herein, the term “sense region” refers to a nucleotide sequenceof a siNA molecule complementary (partially or fully) to an antisenseregion of the siNA molecule. The sense strand of a siNA molecule caninclude a nucleic acid sequence having homology with a target nucleicacid sequence. As used herein, “sense strand” refers to nucleic acidmolecule that includes a sense region and may also include additionalnucleotides.

As used herein, the term “antisense region” refers to a nucleotidesequence of a siNA molecule complementary (partially or fully) to atarget nucleic acid sequence. The antisense strand of a siNA moleculecan optionally include a nucleic acid sequence complementary to a senseregion of the siNA molecule. As used herein, “antisense strand” refersto nucleic acid molecule that includes an antisense region and may alsoinclude additional nucleotides.

As used herein, the term “RNA” refers to a molecule that includes atleast one ribonucleotide residue.

As used herein, the term “duplex region” refers to the region in twocomplementary or substantially complementary oligonucleotides that formbase pairs with one another, either by Watson-Crick base pairing or anyother manner that allows for a duplex between oligonucleotide strandsthat are complementary or substantially complementary. For example, anoligonucleotide strand having 21 nucleotide units can base pair withanother oligonucleotide of 21 nucleotide units, yet only 19 bases oneach strand are complementary or substantially complementary, such thatthe “duplex region” consists of 19 base pairs. The remaining base pairsmay, for example, exist as 5′ and 3′ overhangs. Further, within theduplex region, 100% complementarity is not required; substantialcomplementarity is allowable within a duplex region. Substantialcomplementarity refers to complementarity between the strands such thatthey are capable of annealing under biological conditions. Techniques toempirically determine if two strands are capable of annealing underbiological conditions are well know in the art. Alternatively, twostrands can be synthesized and added together under biologicalconditions to determine if they anneal to one another.

As used herein, the terms “non-pairing nucleotide analog” means anucleotide analog which includes a non-base pairing moiety including butnot limited to: 6 des amino adenosine (Nebularine), 4-Me-indole,3-nitropyrrole, 5-nitroindole, Ds, Pa, N3-Me ribo U, N3-Me riboT, N3-MedC, N3-Me-dT, N1-Me-dG, N1-Me-dA, N3-ethyl-dC, N3-Me dC. In someembodiments the non-base pairing nucleotide analog is a ribonucleotide.In other embodiments it is a deoxyribonucleotide.

As used herein, the term, “terminal functional group” includes withoutlimitation a halogen, alcohol, amine, carboxylic, ester, amide,aldehyde, ketone, ether groups.

An “abasic nucleotide” or “abasic nucleotide analog” is as used hereinmay also be often referred to herein and in the art as apseudo-nucleotide or an unconventional moiety. While a nucleotide is amonomeric unit of nucleic acid, generally consisting of a ribose ordeoxyribose sugar, a phosphate, and a base (adenine, guanine, thymine,or cytosine in DNA; adenine, guanine, uracil, or cytosine in RNA). anabasic or pseudo-nucleotide lacks a base, and thus is not strictly anucleotide as the term is generally used in the art. Abasic deoxyribosemoieties include for example, abasic deoxyribosc-3′-phosphate;1,2-dideoxy-D-ribofuranose-3-phosphate;1,4-anhydro-2-deoxy-D-ribitol-3-phosphate. Inverted abasic deoxyribosemoieties include inverted deoxyriboabasic; 3′,5′ inverted deoxyabasic5′-phosphate.

The term “capping moiety” (z″) as used herein includes a moiety whichcan be covalently linked to the 5′ terminus of (N′)y and includes abasicribose moiety, abasic deoxyribose moiety, modifications abasic riboseand abasic deoxyribose moieties including 2′O alkyl modifications;inverted abasic ribose and abasic deoxyribose moieties and modificationsthereof; C6-imino-Pi; a mirror nucleotide including L-DNA and L-RNA;5′OMe nucleotide; and nucleotide analogs including 4′,5′-methylenenucleotide; 1-(β-D-erythrofuranosyl)nucleotide; 4′-thio nucleotide,carbocyclic nucleotide; 5′-amino-alkyl phosphate; 1,3-diamino-2-propylphosphate, 3-aminopropyl phosphate; 6-aminohexyl phosphate;12-aminododecyl phosphate; hydroxypropyl phosphate; 1,5-anhydrohexitolnucleotide; alpha-nucleotide; threo-pentofuranosyl nucleotide; acyclic3′,4′-seco nucleotide; 3,4-dihydroxybutyl nucleotide;3,5-dihydroxypentyl nucleotide, 5′-5′-inverted abasic moiety;1,4-butanediol phosphate; 5′-amino; and bridging or non bridgingmethylphosphonate and 5′-mercapto moieties.

Certain capping moieties may be abasic ribose or abasic deoxyribosemoieties; inverted abasic ribose or abasic deoxyribose moieties;C6-amino-Pi; a mirror nucleotide including L-DNA and L-RNA. The nucleicacid molecules as disclosed herein may be synthesized using one or moreinverted nucleotides, for example inverted thymidine or inverted adenine(for example see Takei, et al., 2002. JBC 277(26)23800-06).

The term “unconventional moiety” as used herein refers to non-nucleotidemoieties including an abasic moiety, an inverted abasic moiety, ahydrocarbon (alkyl) moiety and derivatives thereof, and further includesa deoxyribonucleotide, a modified deoxyribonucleotide, a mirrornucleotide (L-DNA or L-RNA), a non-base pairing nucleotide analog and anucleotide joined to an adjacent nucleotide by a 2′-5′ internucleotidephosphate bond; bridged nucleic acids including LNA and ethylene bridgednucleic acids, linkage modified (e.g. PACE) and base modifiednucleotides as well as additional moieties explicitly disclosed hereinas unconventional moieties.

As used herein, the term “inhibit”, “down-regulate”, or “reduce” withrespect to gene expression means the expression of the gene, or level ofRNA molecules or equivalent RNA molecules encoding one or more proteinsor protein subunits (e.g., mRNA), or activity of one or more proteins orprotein subunits, is reduced below that observed in the absence of aninhibitory factor (such as a nucleic acid molecule, e.g., an siNA, forexample having structural features as described herein); for example theexpression may be reduced to 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%,10%, 5% or less than that observed in the absence of an inhibitor.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a bar graph showing the effect of GFP siNA on various reportercell lines. Cell lines were established by lenti-viral induction ofhuman HSP47 cDNA-GFP or rat GP46 cDNA-GFP construct into HEK293, humanfibrosarcoma cell line HT1080, human HSC line hTERT or NRK cell line.Negative control siNA or siNA against GFP was introduced into the cellsand GFP fluorescence was measured. The results showed that siNA againstGFP knocks down the fluorescence to different extent in different celllines. 293 HSP47-GFP and 293_GP46-GFP cell lines were selected forsiHsp47 screening due to their easiness of being transfected andsensitivity to fluorescence knockdown.

FIG. 2 is a series of bar graphs showing the cytotoxicity and knockdownefficiency of various siHsp47s in 293_HSP47-GFP and 293_GP46-GFP celllines. The result showed that siHsp47-C, siHsp47-2 and siHsp47-2defficiently knockdown both human HSP47 and rat GP46 (the human hsp47homolog) without substantial cytotoxicity. siGp46A against GP46 does notknock down human HSP47. Additionally, the newly designed siHsp47soutperformed siGp46A in knocking down rat GP46.

FIG. 3 is a bar graph showing the knock down effect of various siHsp47son hsp47 mRNA, measured by TaqMan® qPCR using the human HSC cell linehTERT. The Y axis represents the remaining mRNA level of hsp47. HSP47-Cwas most effective among all the hsp47 siNAs tested.

FIG. 4 is a bar graph showing the effect of different hsp47 siNAs oncollagen I expression in hTERT cells. The level of collagen I mRNAlevels were measured by real-time quantitative PCR using TaqMan® probe.The Y axis represents the remaining mRNA expression level of collagen I.The result showed that collagen I mRNA level is significantly reduced inthe cells treated with some of the candidates (siHsp47-2, siHsp47-2d,and their combination with siHsp47-1).

FIG. 5 shows the decrease in fibrotic areas of the liver in animalstreated with siHSP47.

FIG. 6 is an exemplary nucleic acid sequence of human hsp47 mRNA cDNA(SEQ ID NO: 1; based on the cDNA disclosed in GenBank accession number:NM 001235).

FIG. 7 is an exemplary amino acid sequence of human hsp47 (SEQ ID NO:2).

FIG. 8 is protein coding nucleic acid sequence of human hsp47 cDNA (SEQID NO:59), which corresponds to nucleotides 230-1486 of SEQ ID NO: 1.

FIGS. 9A-91 show plasma stability of Compound_(—)1, Compound_(—)2,Compound 3, Compound_(—)4, Compound_(—)5, Compound_(—)6, Compound_(—)7,Compound_(—)8 and Compound_(—)9, respectively, as detected by ethidiumbromide staining.

FIGS. 10A-10I show ontarget/off-target activity of Compound_(—)1,Compound_(—)2, Compound_(—)3, Compound_(—)4, Compound_(—)5,Compound_(—)6, Compound_(—)7, Compound_(—)8 and Compound_(—)9,respectively. AS_CM shows activity of antisense strand of compound to aplasmid comprising a full match insert; AS_SM shows activity ofantisense strand of compound to a plasmid comprising seed sequenceinsert; S_CM shows activity of sense strand of compound to a plasmidcomprising a full match insert. All assays were performed in humancells, except for data shown in FIG. 10F which was performed in ratREF52 cells.

DETAILED DESCRIPTION OF THE INVENTION RNA Interference and siNA NucleicAcid Molecules

RNA interference refers to the process of sequence-specificpost-transcriptional gene silencing in animals mediated by shortinterfering RNAs (siRNAs) (Zamore et al., 2000, Cell, 101, 25-33; Fireet al., 1998, Nature, 391, 806; Hamilton et al., 1999, Science, 286,950-951; Lin et al., 1999, Nature, 402, 128-129; Sharp, 1999, Genes &Dev., 13:139-141; and Strauss, 1999, Science, 286, 886). Thecorresponding process in plants (Heifetz et al., International PCTPublication No. WO 99/61631) is often referred to aspost-transcriptional gene silencing (PTGS) or RNA silencing. The processof post-transcriptional gene silencing is thought to be anevolutionarily-conserved cellular defense mechanism used to prevent theexpression of foreign genes (Fire et al., 1999, Trends Genet., 15, 358).Such protection from foreign gene expression may have evolved inresponse to the production of double-stranded RNAs (dsRNAs) derived fromviral infection or from the random integration of transposon elementsinto a host genome via a cellular response that specifically destroyshomologous single-stranded RNA or viral genomic RNA. The presence ofdsRNA in cells triggers the RNAi response through a mechanism that hasyet to be fully characterized. This mechanism appears to be differentfrom other known mechanisms involving double stranded RNA-specificribonucleases, such as the interferon response that results fromdsRNA-mediated activation of protein kinase PKR and 2′,5′-oligoadenylatesynthetase resulting in non-specific cleavage of mRNA by ribonuclease L(see for example U.S. Pat. Nos. 6,107,094; 5,898,031; Clemens et al.,1997, J. Interferon & Cytokine Res., 17, 503-524; Adah et al., 2001,Curr. Med. Chem., 8, 1189).

The presence of long dsRNAs in cells stimulates the activity of aribonuclease III enzyme referred to as dicer (Bass, 2000, Cell, 101,235; Zamore et al., 2000, Cell, 101, 25-33; Hammond et al., 2000,Nature, 404, 293). Dicer is involved in the processing of the dsRNA intoshort pieces of dsRNA known as short interfering RNAs (siRNAs) (Zamoreet al., 2000, Cell, 101, 25-33; Bass, 2000, Cell, 101, 235; Berstein etal., 2001, Nature, 409, 363). Short interfering RNAs derived from diceractivity are typically about 21 to about 23 nucleotides in length andinclude about 19 base pair duplexes (Zamore et al., 2000, Cell, 101,25-33; Elbashir et al., 2001, Genes Dev., 15, 188). Dicer has also beenimplicated in the excision of 21- and 22-nucleotide small temporal RNAs(stRNAs) from precursor RNA of conserved structure that are implicatedin translational control (Hutvagner et al., 2001, Science, 293, 834).The RNAi response also features an endonuclease complex, commonlyreferred to as an RNA-induced silencing complex (RISC), which mediatescleavage of single-stranded RNA having sequence complementary to theantisense strand of the si RNA duplex. Cleavage of the target RNA takesplace in the middle of the region complementary to the antisense strandof the siRNA duplex (Elbashir et al., 2001, Genes Dev., 15, 188).

RNAi has been studied in a variety of systems. Fire et al., 1998,Nature, 391, 806, were the first to observe RNAi in C. elegans.Bahramian and Zarbl, 1999, Molecular and Cellular Biology, 19, 274-283and Wianny and Goetz, 1999, Nature Cell Biol., 2, 70, describe RNAimediated by dsRNA in mammalian systems. Hammond et al., 2000, Nature,404, 293, describe RNAi in Drosophila cells transfected with dsRNA.Elbashir et al., 2001, Nature, 411, 494 and Tuschl et al., InternationalPCT Publication No. WO 01/75164, describe RNAi induced by introductionof duplexes of synthetic 21-nucleotide RNAs in cultured mammalian cellsincluding human embryonic kidney and HeLa cells. Recent work inDrosophila embryonic lysates (Elbashir et al., 2001, EMBO J., 20, 6877and Tuschl et al., International PCT Publication No. WO 01/75164) hasrevealed certain requirements for siRNA length, structure, chemicalcomposition, and sequence that are essential to mediate efficient RNAiactivity.

Nucleic acid molecules (for example having structural features asdisclosed herein) may inhibit or down regulate gene expression or viralreplication by mediating RNA interference “RNAi” or gene silencing in asequence-specific manner; see e.g., Zamore et al., 2000, Cell, 101,25-33; Bass, 2001, Nature, 411, 428-429; Elbashir et al., 2001, Nature,411, 494-498; and Kreutzer et al., International PCT Publication No. WO00/44895; Zernicka-Goetz et al., International PCT Publication No. WO01/36646; Fire, International PCT Publication No. WO 99/32619; Plaetincket al., International PCT Publication No. WO 00/01846; Mello and Fire,International PCT Publication No. WO 01/29058; Deschamps-Depaillette,International PCT Publication No. WO 99/07409; and Li et al.,International PCT Publication No. WO 00/44914; Allshire, 2002, Science,297, 1818-1819; Volpe et al., 2002, Science, 297, 1833-1837; Jenuwein,2002, Science, 297, 2215-2218; and Hall et al., 2002, Science, 297,2232-2237; Hutvagner and Zamore, 2002, Science, 297, 2056-60; McManus etal., 2002, RNA, 8, 842-850; Reinhart et al., 2002, Gene & Dev., 16,1616-1626; and Reinhart & Bartel, 2002, Science, 297, 1831).

An siNA nucleic acid molecule can be assembled from two separatepolynucleotide strands, where one strand is the sense strand and theother is the antisense strand in which the antisense and sense strandsare self-complementary (i.e. each strand includes nucleotide sequencethat is complementary to nucleotide sequence in the other strand); suchas where the antisense strand and sense strand form a duplex or doublestranded structure having any length and structure as described hereinfor nucleic acid molecules as provided, for example wherein the doublestranded region (duplex region) is about 15 to about 49 (e.g., about 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35,36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, or 49 base pairs);the antisense strand includes nucleotide sequence that is complementaryto nucleotide sequence in a target nucleic acid molecule (i.e., hsp47mRNA) or a portion thereof and the sense strand includes nucleotidesequence corresponding to the target nucleic acid sequence or a portionthereof (e.g., about 17 to about 49 or more nucleotides of the nucleicacid molecules herein are complementary to the target nucleic acid or aportion thereof).

In certain aspects and embodiments a nucleic acid molecule (e.g., a siNAmolecule) provided herein may be a “RISC length” molecule or may be aDicer substrate as described in more detail below.

An siNA nucleic acid molecule may include separate sense and antisensesequences or regions, where the sense and antisense regions arecovalently linked by nucleotide or non-nucleotide linkers molecules asis known in the art, or are alternately non-covalently linked by ionicinteractions, hydrogen bonding, van der Waals interactions, hydrophobicinteractions, and/or stacking interactions. Nucleic acid molecules mayinclude a nucleotide sequence that is complementary to nucleotidesequence of a target gene. Nucleic acid molecules may interact withnucleotide sequence of a target gene in a manner that causes inhibitionof expression of the target gene.

Alternatively, an siNA nucleic acid molecule is assembled from a singlepolynucleotide, where the self-complementary sense and antisense regionsof the nucleic acid molecules are linked by means of a nucleic acidbased or non-nucleic acid-based linker(s), i.e., the antisense strandand the sense strand are part of one single polynucleotide that havingan antisense region and sense region that fold to form a duplex region(for example to form a “hairpin” structure as is well known in the art).Such siNA nucleic acid molecules can be a polynucleotide with a duplex,asymmetric duplex, hairpin or asymmetric hairpin secondary structure,having self-complementary sense and antisense regions, wherein theantisense region includes nucleotide sequence that is complementary tonucleotide sequence in a separate target nucleic acid molecule or aportion thereof and the sense region having nucleotide sequencecorresponding to the target nucleic acid sequence (e.g., a sequence ofhsp47 mRNA). Such siNA nucleic acid molecules can be a circularsingle-stranded polynucleotide having two or more loop structures and astem comprising self-complementary sense and antisense regions, whereinthe antisense region includes nucleotide sequence that is complementaryto nucleotide sequence in a target nucleic acid molecule or a portionthereof and the sense region having nucleotide sequence corresponding tothe target nucleic acid sequence or a portion thereof, and wherein thecircular polynucleotide can be processed either in vivo or in vitro togenerate an active nucleic acid molecule capable of mediating RNAi.

The following nomenclature is often used in the art to describe lengthsand overhangs of siNA molecules and may be used throughout thespecification and Examples. Names given to duplexes indicate the lengthof the oligomers and the presence or absence of overhangs. For example,a “21+2” duplex contains two nucleic acid strands both of which are 21nucleotides in length, also termed a 21-mer siRNA duplex or a 21-mernucleic acid and having a 2 nucleotides 3′-overhang. A “21−2” designrefers to a 21-mer nucleic acid duplex with a 2 nucleotides 5′-overhang.A 21−0 design is a 21-mer nucleic acid duplex with no overhangs (blunt).A “21+2UU” is a 21-mer duplex with 2-nucleotides 3′-overhang and theterminal 2 nucleotides at the 3′-ends are both U residues (which mayresult in mismatch with target sequence). The aforementionednomenclature can be applied to siNA molecules of various lengths ofstrands, duplexes and overhangs (such as 19−0, 21+2, 27+2, and thelike). In an alternative but similar nomenclature, a “25/27” is anasymmetric duplex having a 25 base sense strand and a 27 base antisensestrand with a 2- nucleotides 3′-overhang. A “27/25” is an asymmetricduplex having a 27 base sense strand and a 25 base antisense strand.

Chemical Modifications

In certain aspects and embodiments, nucleic acid molecules (e.g., siNAmolecules) as provided herein include one or more modifications (orchemical modifications). In certain embodiments, such modificationsinclude any changes to a nucleic acid molecule or polynucleotide thatwould make the molecule different than a standard ribonucleotide or RNAmolecule (i.e., that includes standard adenosine, cytosine, uracil, orguanosine moieties); which may be referred to as an “unmodified”ribonucleotide or unmodified ribonucleic acid. Traditional DNA bases andpolynucleotides having a 2′-deoxy sugar represented by adenosine,cytosine, thymine, or guanosine moieties may be referred to as an“unmodified deoxyribonucleotide” or “unmodified deoxyribonucleic acid”;accordingly, the term “unmodified nucleotide” or “unmodified nucleicacid” as used herein refers to an “unmodified ribonucleotide” or“unmodified ribonucleic acid” unless there is a clear indication to thecontrary. Such modifications can be in the nucleotide sugar, nucleotidebase. nucleotide phosphate group and/or the phosphate backbone of apolynucleotide.

In certain embodiments modifications as disclosed herein may be used toincrease RNAi activity of a molecule and/or to increase the in vivostability of the molecules, particularly the stability in serum, and/orto increase bioavailability of the molecules. Non-limiting examples ofmodifications include without limitation internucleotide orinternucleoside linkages; deoxynucleotides or dideoxyribonucleotides atany position and strand of the nucleic acid molecule; nucleic acid(e.g., ribonucleic acid) with a modification at the 2′-positionpreferably selected from an amino, fluoro, methoxy, alkoxy and alkyl;2′-deoxyribonucleotides, 2′-O-methyl ribonucleotides, 2′-deoxy-2′-fluororibonucleotides, “universal base” nucleotides, “acyclic” nucleotides,5-C-methyl nucleotides, biotin group, and terminal glyceryl and/orinverted deoxy abasic residue incorporation, sterically hinderedmolecules, such as fluorescent molecules and the like. Other nucleotidesmodifiers could include 3′-deoxyadenosine (cordycepin),3′-azido-3′-deoxythymidine (AZT), 2′,3′-dideoxyinosine (ddI),2′,3′-dideoxy-3′-thiacytidine (3TC),2′,3′-didehydro-2′,3′-dideoxythymidine (d4T) and the monophosphatenucleotides of 3′-azido-3′-deoxythymidine (AZT),2′,3′-dideoxy-3′-thiacytidine (3TC) and2′,3′-didehydro-2′,3′-dide-oxythymidine (d4T). Further details onvarious modifications are described in more detail below.

Modified nucleotides include those having a Northern conformation (e.g.,Northern pseudorotation cycle, see for example Saenger, Principles ofNucleic Acid Structure, Springer-Verlag ed., 1984). Non-limitingexamples of nucleotides having a northern configuration include lockednucleic acid (LNA) nucleotides (e.g., 2′-O,4′-C-methylene-(D-ribofuranosyl) nucleotides); 2′-methoxyethoxy (MOE)nucleotides; 2′-methyl-thio-ethyl, 2′-deoxy-2′-fluoro nucleotides,2′-deoxy-2′-chloro nucleotides, 2′-azido nucleotides, and 2′-O-methylnucleotides. Locked nucleic acids, or LNA's are described, for example,in Elman et al., 2005; Kurreck et al., 2002; Crinelli et al., 2002;Braasch and Corey, 2001; Bondensgaard et al., 2000; Wahlestedt et al.,2000; and International Patent Publication Nos. WO 00/47599, WO99/14226, and WO 98/39352 and WO 2004/083430. In one embodiment, an LNAis incorporated at the 5′ terminus of the sense strand.

Chemical modifications also include unlocked nucleic acids, or UNAs,which are non-nucleotide, acyclic analogues, in which the C2′-C3′ bondis not present (although UNAs are not truly nucleotides, they areexpressly included in the scope of “modified” nucleotides or modifiednucleic acids as contemplated herein). In particular embodiments,nucleic acid molecules with an overhang may be modified to have UNAs atthe overhang positions (i.e., 2 nucleotide overhand). In otherembodiments, UNAs are included at the 3′- or 5′-ends. A UNA may belocated anywhere along a nucleic acid strand, i.e. in position 7.Nucleic acid molecules may contain one or more than UNA. Exemplary UNAsare disclosed in Nucleic Acids Symposium Series No. 52 p. 133-134(2008). In certain embodiments a nucleic acid molecule (e.g., a siNAmolecule) as described herein include one or more UNAs; or one UNA. Insome embodiments, a nucleic acid molecule (e.g., a siNA molecule) asdescribed herein that has a 3′-overhang include one or two UNAs in the3′ overhang. In some embodiments a nucleic acid molecule (e.g., a siNAmolecule) as described herein includes a UNA (for example one UNA) inthe antisense strand; for example in position 6 or position 7 of theantisense strand. Chemical modifications also include non-pairingnucleotide analogs, for example as disclosed herein. Chemicalmodifications further include unconventional moieties as disclosedherein.

Chemical modifications also include terminal modifications on the 5′and/or 3′ part of the oligonucleotides and are also known as cappingmoieties. Such terminal modifications are selected from a nucleotide, amodified nucleotide, a lipid, a peptide, and a sugar.

Chemical modifications also include six membered “six membered ringnucleotide analogs.” Examples of six-membered ring nucleotide analogsare disclosed in Allart, et al (Nucleosides & Nucleotides, 1998,17:1523-1526,; and Perez-Perez, et al., 1996, Bioorg. and Medicinal ChemLetters 6:1457-1460) Oligonucleotides including 6-membered ringnucleotide analogs including hexitol and altritol nucleotide monomersare disclosed in International patent application publication No. WO2006/047842.

Chemical modifications also include “mirror” nucleotides which have areversed chirality as compared to normal naturally occurring nucleotide;that is a mirror nucleotide may be an “L-nucleotide” analogue ofnaturally occurring D-nucleotide (see U.S. Pat. No. 6,602,858). Mirrornucleotides may further include at least one sugar or base modificationand/or a backbone modification, for example, as described herein, suchas a phosphorothioate or phosphonate moiety. U.S. Pat. No. 6,602,858discloses nucleic acid catalysts including at least one L-nucleotidesubstitution. Mirror nucleotides include for example L-DNA(L-deoxyriboadenosine-3′-phosphate (mirror dA);L-deoxyribocytidine-3′-phosphate (mirror dC);L-deoxyriboguanosine-3′-phosphate (mirror dG);L-deoxyribothymidine-3′-phosphate (mirror image dT)) and L-RNA(L-riboadenosine-3′-phosphate (mirror rA); L-ribocytidine-3-phosphate(mirror rC); L-riboguanosine-3′-phosphate (mirror rG);L-ribouracil-3′-phosphate (mirror dU).

In some embodiments, modified ribonucleotides include modifieddeoxyribonucleotides, for example 5′OMe DNA(5-methyl-deoxyriboguanosine-3′-phosphate) which may be useful as anucleotide in the 5′ terminal position (position number 1); PACE(deoxyriboadenosine 3′ phosphonoacetate, deoxyribocytidine 3′phosphonoacetate, deoxyriboguanosine 3′ phosphonoacetate,deoxyribothymidine 3′ phosphonoacetate.

Modifications may be present in one or more strands of a nucleic acidmolecule disclosed herein, e.g., in the sense strand, the antisensestrand, or both strands. In certain embodiments, the antisense strandmay include modifications and the sense strand my only includeunmodified RNA.

Nucleobases

Nucleobases of the nucleic acid disclosed herein may include unmodifiedribonucleotides (purines and pyrimidines) such as adenine, guanine,cytosine, uracil. The nucleobases in one or both strands can be modifiedwith natural and synthetic nucleobases such as, thymine, xanthine,hypoxanthine, inosine, 2-aminoadenine, 6-methyl and other alkylderivatives of adenine and guanine, any “universal base” nucleotides;2-propyl and other alkyl derivatives of adenine and guanine,5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil,cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo,amino, thiol, thioalkyl, hydroxyl and other 8-substituted adenines andguanines, 5-trifluoromethyl and other 5-substituted uracils andcytosines, 7-methylguanine, deazapurines, heterocyclic substitutedanalogs of purines and pyrimidines, e.g., aminoethyoxy phenoxazine,derivatives of purines and pyrimidines (e.g., 1-alkyl-, 1-alkenyl-,heteroaromatic- and 1-alkynyl derivatives) and tautomers thereof,8-oxo-N-6-methyladenine, 7-diazaxanthine, 5-methylcytosine,5-methyluracil, 5-(1-propynyl)uracil, 5-(1-propynyl) cytosine and4,4-ethanocytosine). Other examples of suitable bases includenon-purinyl and non-pyrimidinyl bases such as 2-aminopyridine andtriazines.

Sugar Moieties

Sugar moieties in nucleic acid disclosed herein may include2′-hydroxyl-pentofuranosyl sugar moiety without any modification.Alternatively, sugar moieties can be modified such as,2′-deoxy-pentofuranosyl sugar moiety, D-ribose, hexose, modification atthe 2′ position of the pentofuranosyl sugar moiety such as 2′-O-alkyl(including 2′-O-methyl and 2′-O-ethyl), i.e., 2′-alkoxy, 2′-amino,2′-O-allyl, 2′-S-alkyl, 2′-halogen (including 2′-fluoro, chloro, andbromo), 2′-methoxyethoxy, 2′-O-methoxyethyl, 2′-O-2-methoxyethyl,2′-allyloxy (—OCH2CH═CH2), 2′-propargyl, 2′-propyl, ethynyl, ethenyl,propenyl, CF, cyano, imidazole, carboxylate, thioate, C1 to C10 loweralkyl, substituted lower alkyl, alkaryl or aralkyl, OCF3, OCN, O-, S-,or N-alkyl; O-, S, or N-alkenyl; SOCH3; SO2CH3; ONO2; NO2, N3;heterozycloalkyl; heterozycloalkaryl; aminoalkylamino; polyalkylamino orsubstituted silyl, as, among others, for example as described inEuropean patents EP 0 586 520 B1 or EP 0 618 925 B1.

Alkyl group includes saturated aliphatic groups, includingstraight-chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl,hexyl, heptyl, octyl, nonyl, decyl, etc.), branched-chain alkyl groups(isopropyl, tert-butyl, isobutyl, etc.), cycloalkyl(alicyclic) groups(cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl), alkylsubstituted cycloalkyl groups, and cycloalkyl substituted alkyl groups.In certain embodiments, a straight chain or branched chain alkyl has 6or fewer carbon atoms in its backbone (e.g., C1-C6 for straight chain,C3-C6 for branched chain), and more preferably 4 or fewer. Likewise,preferred cycloalkyls may have from 3-8 carbon atoms in their ringstructure, and more preferably have 5 or 6 carbons in the ringstructure. The term C1-C6 includes alkyl groups containing 1 to 6 carbonatoms. The alkyl group can be substituted alkyl group such as alkylmoieties having substituents replacing a hydrogen on one or more carbonsof the hydrocarbon backbone. Such substituents can include, for example,alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy,alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, alkylcarbonyl,arylcarbonyl, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,phosphonato, phosphinato, cyano, amino (including alkyl amino,dialkylamino, arylamino, diarylamino, and alkylarylamino), acylamino(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety.

Alkoxy group includes substituted and unsubstituted alkyl, alkenyl, andalkynyl groups covalently linked to an oxygen atom. Examples of alkoxygroups include methoxy, ethoxy, isopropyloxy, propoxy, butoxy, andpentoxy groups. Examples of substituted alkoxy groups includehalogenated alkoxy groups. The alkoxy groups can be substituted withgroups such as alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkyl aryl amino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,alkylaryl, or an aromatic or heteroaromatic moieties. Examples ofhalogen substituted alkoxy groups include, but are not limited to,fluoromethoxy, difluoromethoxy, trifluoromethoxy, chloromethoxy,dichloromethoxy, trichloromethoxy, etc.

In some embodiments, the pentafuronosyl ring may be replaced withacyclic derivatives lacking the C2′-C3′-bond of the pentafuronosyl ring.For example, acyclonucleotides may substitute a 2-hydroxyethoxymethylgroup for-the 2′-deoxyribofuranosyl sugar normally present in dNMPs.

Halogens include fluorine, bromine, chlorine, iodine.

Backbone

The nucleoside subunits of the nucleic acid disclosed herein may belinked to each other by phosphodiester bond. The phosphodiester bond maybe optionally substituted with other linkages. For example,phosphorothioate, thiophosphate-D-ribose entities, triester, thioate,2′-S′ bridged backbone (may also he referred to as 5′-2′ or 2′5′nucleotide or 2′5′ ribonucleotide), PACE, 3′-(or -5′)deoxy-3′-(or-5′)thio-phosphorothioate, phosphorodithioate, phosphoroselenates,3′-(or -5′)deoxy phosphinates, borano phosphates, 3′-(or-5′)deoxy-3′-(or 5′-)amino phosphoramidates, hydrogen phosphonates,phosphonates, borano phosphate esters, phosphoramidates, alkyl or arylphosphonates and phosphotriester modifications such asalkylphosphotriesters, phosphotriester phosphorus linkages,5′-ethoxyphosphodiester, P-alkyl oxyphosphotriester, methylphosphonate,and nonphosphorus containing linkages for example, carbonate, carbamate,silyl, sulfur, sulfonate, sulfonamide. formacetal, thioformacetyl,oxime, methyleneimino, methylenemethylimino, methylenehydrazo,methylenedimethylhydrazo and methyleneoxymethylimino linkages.

Nucleic acid molecules disclosed herein may include a peptide nucleicacid (PNA) backbone. The PNA backbone is includes repeatingN-(2-aminoethyl)-glycine units linked by peptide bonds. The variousbases such as purine, pyrimidine, natural and synthetic bases are linkedto the backbone by methylene carbonyl bonds.

Terminal Phosphates

Modifications can be made at terminal phosphate groups. Non-limitingexamples of different stabilization chemistries can be used, e.g., tostabilize the 3′-end of nucleic acid sequences, including (1)[3-3′]-inverted deoxyribose; (2) deoxyribonucleotide; (3)[5′-3′]-3′-deoxyribonucleotide; (4) [5′-3′]-ribonucleotide; (5)[5′-3′]-3′-O-methyl ribonucleotide; (6) 3′-glyceryl; (7)[3′-5′]-3′-deoxyribonucleotide; (8) [3′-3′]-deoxyribonucleotide; (9)[5′-2′]-deoxyribonucleotide; and (10) [5-3′]-dideoxyribonucleotide. Inaddition to unmodified backbone chemistries can be combined with one ormore different backbone modifications described herein.

Exemplary chemically modified terminal phosphate groups include thoseshown below:

Conjugates

Modified nucleotides and nucleic acid molecules (e.g., siNA molecules)as provided herein may include conjugates, for example, a conjugatecovalently attached to the chemically-modified nucleic acid molecule.Non-limiting examples of conjugates include conjugates and ligandsdescribed in Vargeese et al., U.S. Ser. No. 10/427,160. The conjugatemay be covalently attached to a nucleic acid molecule (such as an siNAmolecule) via a biodegradable linker. The conjugate molecule may beattached at the 3′-end of either the sense strand, the antisense strand,or both strands of the chemically-modified nucleic acid molecule. Theconjugate molecule may be attached at the 5′-end of either the sensestrand, the antisense strand, or both strands of the chemically-modifiednucleic acid molecule. The conjugate molecule may be attached both the3′-end and 5′-end of either the sense strand, the antisense strand, orboth strands of the chemically-modified nucleic acid molecule, or anycombination thereof. In one embodiment, a conjugate molecule may includea molecule that facilitates delivery of a chemically-modified nucleicacid molecule into a biological system, such as a cell. In anotherembodiment, the conjugate molecule attached to the chemically-modifiednucleic acid molecule is a polyethylene glycol, human serum albumin, ora ligand for a cellular receptor that can mediate cellular uptake.Examples of specific conjugate molecules contemplated by the instantinvention that can be attached to chemically-modified nucleic acidmolecules are described in Vargeese et al., U.S. Ser. No. 10/201,394.

Linkers

A nucleic acid molecule provided herein (e.g., an siNA) molecule mayinclude a nucleotide, non-nucleotide, or mixed nucleotide/non-nucleotidelinker that joins the sense region of the nucleic acid to the antisenseregion of the nucleic acid. A nucleotide linker can be a linker of ≧2nucleotides in length, for example about 3, 4, 5, 6, 7, 8, 9, or 10nucleotides in length. The nucleotide linker can be a nucleic acidaptamer. By “aptamer” or “nucleic acid aptamer” as used herein refers toa nucleic acid molecule that binds specifically to a target moleculewherein the nucleic acid molecule has sequence that includes a sequencerecognized by the target molecule in its natural setting. Alternately,an aptamer can be a nucleic acid molecule that binds to a targetmolecule (such as hsp47 mRNA) where the target molecule does notnaturally bind to a nucleic acid. For example, the aptamer can be usedto bind to a ligand-binding domain of a protein, thereby preventinginteraction of the naturally occurring ligand with the protein. This isa non-limiting example and those in the art will recognize that otherembodiments can be readily generated using techniques generally known inthe art. See e.g., Gold et al.; 1995, Annu. Rev. Biochem., 64, 763;Brody and Gold, 2000, J. Biotechnol., 74, 5; Sun, 2000, Curr. Opin. Mol.Ther., 2, 100; Kusser, 2000, J. Biotechnol., 74, 27; Hermann and Patel,2000, Science, 287, 820; and Jayasena, 1999, Clinical Chemistry, 45,1628.

A non-nucleotide linker may include an abasic nucleotide, polyether,polyamine, polyamide, peptide, carbohydrate, lipid, polyhydrocarbon, orother polymeric compounds (e.g. polyethylene glycols such as thosehaving between 2 and 100 ethylene glycol units). Specific examplesinclude those described by Seela and Kaiser, Nucleic Acids Res. 1990,18:6353 and Nucleic Acids Res. 1987, 15:3113; Cload and Schepartz, J.Am. Chem. Soc. 1991, 113:6324; Richardson and Schepartz, J. Am. Chem.Soc. 1991, 113:5109; Ma et al., Nucleic Acids Res. 1993, 21:2585 andBiochemistry 1993, 32:1751; Durand et al., Nucleic Acids Res. 1990,18:6353; McCurdy et al., Nucleosides & Nucleotides 1991, 10:287; Jschkeet al., Tetrahedron Lett. 1993, 34:301; Ono et al., Biochemistry 1991,30:9914; Arnold et al., International Publication No. WO 89/02439; Usmanet al., International Publication No. WO 95/06731; Dudycz et al.,International Publication No. WO 95/11910 and Ferentz and Verdine, J.Am. Chem. Soc. 1991, 113:4000.

5′ Ends, 3′ Ends and Overhangs

Nucleic acid molecules disclosed herein (e.g., siNA molecules) may beblunt-ended on both sides, have overhangs on both sides or a combinationof blunt and overhang ends. Overhangs may occur on either the 5′- or3′-end of the sense or antisense strand.

5′- and/or 3′-ends of double stranded nucleic acid molecules (e.g.,siNA) may be blunt ended or have an overhang. The 5′-end may be bluntended and the 3′-end has an overhang in either the sense strand or theantisense strand. In other embodiments, the 3′-end may be blunt endedand the 5′-end has an overhang in either the sense strand or theantisense strand. In yet other embodiments, both the 5′- and 3′-end areblunt ended or both the 5′- and 3′-ends have overhangs.

The 5′- and/or 3′-end of one or both strands of the nucleic acid mayinclude a free hydroxyl group. The 5′- and/or 3′-end of any nucleic acidmolecule strand may be modified to include a chemical modification. Suchmodification may stabilize nucleic acid molecules, e.g., the 3′-end mayhave increased stability due to the presence of the nucleic acidmolecule modification. Examples of end modifications (e.g., terminalcaps) include, but are not limited to, abasic, deoxy abasic, inverted(deoxy) abasic, glyceryl, dinucleotide, acyclic nucleotide, amino,fluoro, chloro, bromo, CN, CF, methoxy, imidazole, carboxylate, thioate,C1 to C10 lower alkyl, substituted lower alkyl, alkaryl or aralkyl,OCF3, OCN, O-, S-, or N-alkyl; O-, S-, or N-alkenyl; SOCH3; SO2CH3;ONO2; NO2, N3; heterocycloalkyl; heterocycloalkaryl; aminoalkylamino;polyalkylamino or substituted silyl, as, among others, described inEuropean patents EP 586,520 and EP 618,925 and other modificationsdisclosed herein.

Nucleic acid molecules include those with blunt ends, i.e., ends that donot include any overhanging nucleotides. A nucleic acid molecule caninclude one or more blunt ends. The blunt ended nucleic acid moleculehas a number of base pairs equal to the number of nucleotides present ineach strand of the nucleic acid molecule. The nucleic acid molecule caninclude one blunt end, for example where the 5′-end of the antisensestrand and the 3′-end of the sense strand do not have any overhangingnucleotides. Nucleic acid molecule may include one blunt end, forexample where the 3′-end of the antisense strand and the 5′-end of thesense strand do not have any overhanging nucleotides. A nucleic acidmolecule may include two blunt ends, for example where the 3′-end of theantisense strand and the 5′-end of the sense strand as well as the5′-end of the antisense strand and 3′-end of the sense strand do nothave any overhanging nucleotides. Other nucleotides present in a bluntended nucleic acid molecule can include, for example, mismatches,bulges, loops, or wobble base pairs to modulate the activity of thenucleic acid molecule to mediate RNA interference.

In certain embodiments of the nucleic acid molecules (e.g., siNAmolecules) provided herein, at least one end of the molecule has anoverhang of at least one nucleotide (for example 1 to 8 overhangnucleotides). For example, one or both strands of a double strandednucleic acid molecule disclosed herein may have an overhang at the5′-end or at the 3′-end or both. An overhang may be present at either orboth the sense strand and antisense strand of the nucleic acid molecule.The length of the overhang may be as little as one nucleotide and aslong as 1 to 8 or more nucleotides (e.g., 1, 2, 3, 4, 5, 6, 7 or 8nucleotides; in some preferred embodiments an overhang is 2, 3, 4, 5, 6,7 or 8 nucleotides; for example an overhang may be 2 nucleotides. Thenucleotide(s) forming the overhang may be includedeoxyribonucleotide(s), ribonucleotide(s), natural and non-naturalnucleobases or any nucleotide modified in the sugar, base or phosphategroup such as disclosed herein. A double stranded nucleic acid moleculemay have both 5′- and 3′-overhangs. The overhangs at the 5′- and 3′-endmay be of different lengths. An overhang may include at least onenucleic acid modification which may be deoxyribonucleotide. One or moredeoxyribonucleotides may be at the 5′-terminal. The 3′-end of therespective counter-strand of the nucleic acid molecule may not have anoverhang, more preferably not a deoxyribonucleotide overhang. The one ormore deoxyribonucleotide may be at the 3′-terminal. The 5′-end of therespective counter-strand of the dsRNA may not have an overhang, morepreferably not a deoxyribonucleotide overhang. The overhang in eitherthe 5′- or the 3′-end of a strand may be 1 to 8 (e.g., about 1, 2, 3, 4,5, 6, 7 or 8) unpaired nucleotides, preferably, the overhang is 2-3unpaired nucleotides; more preferably 2 unpaired nucleotides. Nucleicacid molecules may include duplex nucleic acid molecules withoverhanging ends of about 1 to about 20 (e.g., about 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 1, 15, 16, 17, 18, 19 or 20); preferably 1-8(e.g., about 1, 2, 3, 4, 5, 6, 7 or 8) nucleotides, for example, about21-nucleotide duplexes with about 19 base pairs and 3′-terminalmononucleotide, dinucleotide, or trinucleotide overhangs. Nucleic acidmolecules herein may include duplex nucleic acid molecules with bluntends, where both ends are blunt, or alternatively, where one of the endsis blunt. Nucleic acid molecules disclosed herein can include one ormore blunt ends, i.e. where a blunt end does not have any overhangingnucleotides. In one embodiment, the blunt ended nucleic acid moleculehas a number of base pairs equal to the number of nucleotides present ineach strand of the nucleic acid molecule. The nucleic acid molecule mayinclude one blunt end, for example where the 5′-end of the antisensestrand and the 3′-end of the sense strand do not have any overhangingnucleotides. The nucleic acid molecule may include one blunt end, forexample where the 3′-end of the antisense strand and the 5′-end of thesense strand do not have any overhanging nucleotides. A nucleic acidmolecule may include two blunt ends, for example where the 3′-end of theantisense strand and the 5′-end of the sense strand as well as the5′-end of the antisense strand and 3′-end of the sense strand do nothave any overhanging nucleotides. In certain preferred embodiments thenucleic acid compounds are blunt ended. Other nucleotides present in ablunt ended siNA molecule can include, for example, mismatches, bulges,loops, or wobble base pairs to modulate the activity of the nucleic acidmolecule to mediate RNA interference.

In many embodiments one or more, or all, of the overhang nucleotides ofa nucleic acid molecule (e.g., a siNA molecule) as described hereinincludes are modified such as described herein; for example one or more,or all, of the nucleotides may be 2′-deoxynucleotides.

Amount, Location and Patterns of Modifications.

Nucleic acid molecules (e.g., siNA molecules) disclosed herein mayinclude modified nucleotides as a percentage of the total number ofnucleotides present in the nucleic acid molecule. As such, a nucleicacid molecule may include about 5% to about 100% modified nucleotides(e.g., about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% modified nucleotides). Theactual percentage of modified nucleotides present in a given nucleicacid molecule will depend on the total number of nucleotides present inthe nucleic acid. If the nucleic acid molecule is single stranded, thepercent modification can be based upon the total number of nucleotidespresent in the single stranded nucleic acid molecule. Likewise, if thenucleic acid molecule is double stranded, the percent modification canbe based upon the total number of nucleotides present in the sensestrand, antisense strand, or both the sense and antisense strands.

Nucleic acid molecules disclosed herein may include unmodified RNA as apercentage of the total nucleotides in the nucleic acid molecule. Assuch, a nucleic acid molecule may include about 5% to about 100%modified nucleotides (e.g., about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% of totalnucleotides present in a nucleic acid molecule.

A nucleic acid molecule (e.g., an siNA molecule) may include a sensestrand that includes about 1 to about 5, specifically about 1, 2, 3, 4,or 5 phosphorothioate internucleotide linkages, and/or one or more(e.g., about 1, 2, 3, 4, 5, or more) 2′-deoxy, 2′-O-methyl,2′-deoxy-2′-fluoro, and/or one or more (e.g., about 1, 2, 3, 4, 5, ormore) universal base modified nucleotides, and optionally a terminal capmolecule at the 3-end, the 5′-end, or both of the 3′- and 5′-ends of thesense strand; and wherein the antisense strand includes about 1 to about5 or more, specifically about 1, 2, 3, 4, 5, or more phosphorothioateinternucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5,6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro,and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more)universal base modified nucleotides, and optionally a terminal capmolecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends ofthe antisense strand. A nucleic acid molecule may include about 1, 2, 3,4, 5, 6, 7, 8, 9, 10, or more, pyrimidine nucleotides of the senseand/or antisense nucleic acid strand are chemically-modified with2′-deoxy, 2′-O-methyl and/or 2′-deoxy-2′-fluoro nucleotides, with orwithout about 1 to about 5 or more, for example about 1, 2, 3, 4, 5, ormore phosphorothioate internucleotide linkages and/or a terminal capmolecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends,being present in the same or different strand.

A nucleic acid molecule may include about 1 to about 5 or more(specifically about 1, 2, 3, 4, 5 or more) phosphorothioateinternucleotide linkages in each strand of the nucleic acid molecule.

A nucleic acid molecule may include 2′-5′ internucleotide linkages, forexample at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of oneor both nucleic acid sequence strands. In addition, the 2′-5′internucleotide linkage(s) can be present at various other positionswithin one or both nucleic acid sequence strands, for example, about 1,2, 3, 4, 5, 6, 7, 8, 9, 10, or more including every internucleotidelinkage of a pyrimidine nucleotide in one or both strands of the siNAmolecule can include a 2′-S′ internucleotide linkage, or about 1, 2, 3,4, 5, 6, 7, 8, 9, 10, or more including every internucleotide linkage ofa purine nucleotide in one or both strands of the siNA molecule caninclude a 2′-5′ internucleotide linkage.

A chemically-modified short interfering nucleic acid (siNA) molecule mayinclude an antisense region, wherein any (e.g., one or more or all)pyrimidine nucleotides present in the antisense region are2′-deoxy-2′-fluoro pyrimidine nucleotides (e.g., wherein all pyrimidinenucleotides are 2′-deoxy-2′-fluoro pyrimidine nucleotides or alternatelya plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro pyrimidinenucleotides), and wherein any (e.g., one or more or all) purinenucleotides present in the antisense region are 2′-deoxy purinenucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purinenucleotides or alternately a plurality of purine nucleotides are2′-deoxy purine nucleotides).

A chemically-modified short interfering nucleic acid (siNA) molecule mayinclude an antisense region, wherein any (e.g., one or more or all)pyrimidine nucleotides present in the antisense region are2′-deoxy-2′-fluoro pyrimidine nucleotides (e.g., wherein all pyrimidinenucleotides are 2′-deoxy-2′-fluoro pyrimidine nucleotides or alternatelya plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro pyrimidinenucleotides), and wherein any (e.g., one or more or all) purinenucleotides present in the antisense region are 2′-O-methyl purinenucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl purinenucleotides or alternately a plurality of purine nucleotides are2′-O-methyl purine nucleotides).

A chemically-modified short interfering nucleic acid (siNA) moleculecapable of mediating RNA interference (RNAi) against hsp47 inside a cellor reconstituted in vitro system may include a sense region, wherein oneor more pyrimidine nucleotides present in the sense region are2′-deoxy-2′-fluoro pyrimidine nucleotides (e.g., wherein all pyrimidinenucleotides are 2′-deoxy-2′-fluoro pyrimidine nucleotides or alternatelya plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro pyrimidinenucleotides), and one or more purine nucleotides present in the senseregion are 2′-deoxy purine nucleotides (e.g., wherein all purinenucleotides are 2′-deoxy purine nucleotides or alternately a pluralityof purine nucleotides are 2′-deoxy purine nucleotides), and an antisenseregion, wherein one or more pyrimidine nucleotides present in theantisense region are 2′-deoxy-2′-fluoro pyrimidine nucleotides (e.g.,wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro pyrimidinenucleotides or alternately a plurality of pyrimidine nucleotides are2′-deoxy-2′-fluoro pyrimidine nucleotides), and one or more purinenucleotides present in the antisense region are 2′-O-methyl purinenucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl purinenucleotides or alternately a plurality of purine nucleotides are2′-O-methyl purine nucleotides). The sense region and/or the antisenseregion can have a terminal cap modification, such as any modification,that is optionally present at the 3′-end, the 5′-end, or both of the 3′and 5′-ends of the sense and/or antisense sequence. The sense and/orantisense region can optionally further include a 3′-terminal nucleotideoverhang having about 1 to about 4 (e.g., about 1, 2, 3, or 4)2′-deoxynucleotides. The overhang nucleotides can further include one ormore (e.g., about 1, 2, 3, 4 or more) phosphorothioate,phosphonoacetate, and/or thiophosphonoacetate internucleotide linkages.The purine nucleotides in the sense region may alternatively be2′-O-methyl purine nucleotides (e.g., wherein all purine nucleotides are2′-O-methyl purine nucleotides or alternately a plurality of purinenucleotides are 2′-O-methyl purine nucleotides) and one or more purinenucleotides present in the antisense region are 2′-O-methyl purinenucleotides (e.g., wherein all purine nucleotides are 2′-O-methyl purinenucleotides or alternately a plurality of purine nucleotides are2′-O-methyl purine nucleotides). One or more purine nucleotides in thesense region may alternatively be purine ribonucleotides (e.g., whereinall purine nucleotides are purine ribonucleotides or alternately aplurality of purine nucleotides are purine ribonucleotides) and anypurine nucleotides present in the antisense region are 2′-O-methylpurine nucleotides (e.g., wherein all purine nucleotides are 2′-O-methylpurine nucleotides or alternately a plurality of purine nucleotides are2′-O-methyl purine nucleotides). One or more purine nucleotides in thesense region and/or present in the antisense region may alternativelyselected from the group consisting of 2′-deoxy nucleotides, lockednucleic acid (LNA) nucleotides, 2′-methoxyethyl nucleotides,4′-thionucleotides, and 2′-O-methyl nucleotides (e.g., wherein allpurine nucleotides are selected from the group consisting of 2′-deoxynucleotides, locked nucleic acid (LNA) nucleotides, 2′-methoxyethylnucleotides, 4′-thionucleotides, and 2′-O-methyl nucleotides oralternately a plurality of purine nucleotides are selected from thegroup consisting of 2′-deoxy nucleotides, locked nucleic acid (LNA)nucleotides, 2′-methoxyethyl nucleotides, 4′-thionucleotides, and2′-O-methyl nucleotides).

In some embodiments, a nucleic acid molecule (e.g., a siNA molecule) asdescribed herein includes a modified nucleotide (for example onemodified nucleotide) in the antisense strand; for example in position 6or position 7 of the antisense strand.

Modification Patterns and Alternating Modifications

Nucleic acid molecules (e.g., siNA molecules) provided herein may havepatterns of modified and unmodified nucleic acids. A pattern ofmodification of the nucleotides in a contiguous stretch of nucleotidesmay be a modification contained within a single nucleotide or group ofnucleotides that are covalently linked to each other via standardphosphodiester bonds or, at least partially, through phosphorothioatebonds. Accordingly, a “pattern” as contemplated herein, does notnecessarily need to involve repeating units, although it may. Examplesof modification patterns that may be used in conjunction with thenucleic acid molecules (e.g., siNA molecules) provided herein includethose disclosed in Giese, U.S. Pat. No. 7,452,987. For example, nucleicacid molecules (e.g., siNA molecules) provided herein include thosehaving modification patters such as, similar to, or the same as, thepatterns shown diagrammatically in FIG. 2 of the Giese U.S. Pat. No.7,452,987.

A modified nucleotide or group of modified nucleotides may be at the5′-end or 3′-end of the sense or antisense strand, a flanking nucleotideor group of nucleotides is arrayed on both sides of the modifiednucleotide or group, where the flanking nucleotide or group either isunmodified or does not have the same modification of the precedingnucleotide or group of nucleotides. The flanking nucleotide or group ofnucleotides may, however, have a different modification. This sequenceof modified nucleotide or group of modified nucleotides, respectively,and unmodified or differently modified nucleotide or group of unmodifiedor differently modified nucleotides may be repeated one or more times.

In some patterns, the 5′-terminal nucleotide of a strand is a modifiednucleotide while in other patterns the 5′-terminal nucleotide of astrand is an unmodified nucleotide. In some patterns, the 5′-end of astrand starts with a group of modified nucleotides while in otherpatterns, the 5′-terminal end is an unmodified group of nucleotides.This pattern may be either on the first stretch or the second stretch ofthe nucleic acid molecule or on both.

Modified nucleotides of one strand of the nucleic acid molecule may becomplementary in position to the modified or unmodified nucleotides orgroups of nucleotides of the other strand.

There may be a phase shift between modifications or patterns ofmodifications on one strand relative to the pattern of modification ofthe other strand such that the modification groups do not overlap. Inone instance, the shift is such that the modified group of nucleotidesof the sense strand corresponds to the unmodified group of nucleotidesof the antisense strand and vice versa.

There may be a partial shift of the pattern of modification such thatthe modified groups overlap. The groups of modified nucleotides in anygiven strand may optionally be the same length, but may be of differentlengths. Similarly, groups of unmodified nucleotides in any given strandmay optionally be the same length, or of different lengths.

In some patterns, the second (penultimate) nucleotide at the terminus ofthe strand, is an unmodified nucleotide or the beginning of group ofunmodified nucleotides. Preferably, this unmodified nucleotide orunmodified group of nucleotides is located at the 5′-end of the eitheror both the sense and antisense strands and even more preferably at theterminus of the sense strand. An unmodified nucleotide or unmodifiedgroup of nucleotide may be located at the 5′-end of the sense strand. Ina preferred embodiment the pattern consists of alternating singlemodified and unmodified nucleotides.

In some double stranded nucleic acid molecules include a 2′-O-methylmodified nucleotide and a non-modified nucleotide, preferably anucleotide which is not 2′-O-methyl modified, are incorporated on bothstrands in an alternating fashion, resulting in a pattern of alternating2′-O-methyl modified nucleotides and nucleotides that are eitherunmodified or at least do not include a 2′-O-methyl modification. Incertain embodiments, the same sequence of 2′-O-methyl modification andnon-modification exists on the second strand; in other embodiments thealternating 2′-O-methyl modified nucleotides are only present in thesense strand and are not present in the antisense strand; and in yetother embodiments the alternating 2′-O-methyl modified nucleotides areonly present in the sense strand and are not present in the antisensestrand. In certain embodiments, there is a phase shift between the twostrands such that the 2′-O-methyl modified nucleotide on the firststrand base pairs with a non-modified nucleotide(s) on the second strandand vice versa. This particular arrangement, i.e. base pairing of2′-O-methyl modified and non-modified nucleotide(s) on both strands isparticularly preferred in certain embodiments. In certain embodiments,the pattern of alternating 2′-O-methyl modified nucleotides existsthroughout the entire nucleic acid molecule; or the entire duplexregion. In other embodiments the pattern of alternating 2′-O-methylmodified nucleotides exists only in a portion of the nucleic acid; orthe entire duplex region.

In “phase shift” patterns, it may be preferred if the antisense strandstarts with a 2′-O-methyl modified nucleotide at the 5′ end wherebyconsequently the second nucleotide is non-modified, the third, fifth,seventh and so on nucleotides are thus again 2′-O-methyl modifiedwhereas the second, fourth, sixth, eighth and the like nucleotides arenon-modified nucleotides.

Exemplary Modification Locations and Patterns

While exemplary patterns are provided in more detail below, allpermutations of patterns with of all possible characteristics of thenucleic acid molecules disclosed herein and those known in the art arecontemplated (e.g., characteristics include, but are not limited to,length of sense strand, length of antisense strand, length of duplexregion, length of hangover, whether one or both ends of a doublestranded nucleic acid molecule is blunt or has an overhang, location ofmodified nucleic acid, number of modified nucleic acids, types ofmodifications, whether a double overhang nucleic acid molecule has thesame or different number of nucleotides on the overhang of each side,whether a one or more than one type of modification is used in a nucleicacid molecule, and number of contiguous modified/unmodifiednucleotides). With respect to all detailed examples provided below,while the duplex region is shown to be 19 nucleotides, the nucleic acidmolecules provided herein can have a duplex region ranging from 1 to 49nucleotides in length as each strand of a duplex region canindependently be 17-49 nucleotides in length Exemplary patterns areprovided herein.

Nucleic acid molecules may have a blunt end (when n is 0) on both endsthat include a single or contiguous set of modified nucleic acids. Themodified nucleic acid may be located at any position along either thesense or antisense strand. Nucleic acid molecules may include a group ofabout 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48 or 49 contiguous modifiednucleotides. Modified nucleic acids may make up 1%, 2%, 3%, 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95%, 97%, 98% or 100% of a nucleic acid strand. Modifiednucleic acids of the examples immediately below may be in the sensestrand only, the antisense strand only, or in both the sense andantisense strand.

General nucleic acid patters are shown below where X=sense strandnucleotide in the duplex region; X_(a)=5′-overhang nucleotide in thesense strand; X_(b)=3′-overhang nucleotide in the sense strand;Y=antisense strand nucleotide in the duplex region; Y_(a)=3′-overhangnucleotide in the antisense strand; Y_(b)=5′-overhang nucleotide in theantisense strand; and M=a modified nucleotide in the duplex region. Eacha and b are independently 0 to 8 (e.g., 0, 1, 2, 3, 4, 5, 6, 7 or 8).Each X, Y, a and b are independently modified or unmodified. The senseand antisense strands can are each independently 17-49 nucleotides inlength. The examples provided below have a duplex region of 19nucleotides; however, nucleic acid molecules disclosed herein can have aduplex region anywhere between 17 and 49 nucleotides and where eachstrand is independently between 17 and 49 nucleotides in length.

5′ X_(a)XXXXXXXXXXXXXXXXXXXX_(b) 3′ Y_(b)YYYYYYYYYYYYYYYYYYYY_(a)

Further exemplary nucleic acid molecule patterns are shown below whereX=unmodified sense strand nucleotides; x=an unmodified overhangnucleotide in the sense strand; Y=unmodified antisense strandnucleotides; y=an unmodified overhang nucleotide in the antisensestrand; and M=a modified nucleotide. The sense and antisense strands canare each independently 17-49 nucleotides in length. The examplesprovided below have a duplex region of 19 nucleotides; however, nucleicacid molecules disclosed herein can have a duplex region anywherebetween 17 and 49 nucleotides and where each strand is independentlybetween 17 and 49 nucleotides in length.

5′ M _(n)XXXXXXXXXMXXXXXXXXXM _(n) 3′ M _(n)YYYYYYYYYYYYYYYYYYYM _(n) 5′XXXXXXXXXXXXXXXXXXX 3′ YYYYYYYYYMYYYYYYYYY 5′ XXXXXXXXMMXXXXXXXXX 3′YYYYYYYYYYYYYYYYYYY 5′ XXXXXXXXXXXXXXXXXXX 3′ YYYYYYYYMMYYYYYYYYY 5′XXXXXXXXXMXXXXXXXXX 3′ YYYYYYYYYMYYYYYYYYY 5′ XXXXXMXXXXXXXXXXXXX 3′YYYYYYYYYMYYYYYYYYY 5′ MXXXXXXXXXXXXXXXXXX 3′ YYYYYYYYYYYYMYYYYYY 5′XXXXXXXXXXXXXXXXXXM 3′ YYYYYMYYYYYYYYYYYYY 5′ XXXXXXXXXMXXXXXXXX 3′MYYYYYYYYYYYYYYYYY 5′ XXXXXXXMXXXXXXXXXX 3′ YYYYYYYYYYYYYYYYYM 5′XXXXXXXXXXXXXMXXXX 3′ MYYYYYYYYYYYYYYYYY 5′ MMMMMMMMMMMMMMMMMM 3′MMMMMMMMMMMMMMMMMM

Nucleic acid molecules may have blunt ends on both ends with alternatingmodified nucleic acids. The modified nucleic acids may be located at anyposition along either the sense or antisense strand.

5′ MXMXMXMXMXMXMXMXMXM 3′ YMYMYMYMYMYMYMYMYMY 5′ XMXMXMXMXMXMXMXMXMX 3′MYMYMYMYMYMYMYMYMYM 5′ MMXMMXMMXMMXMMXMMXM 3′ YMMYMMYMMYMMYMMYMMY 5′XMMXMMXMMXMMXMMXMMX 3′ MMYMMYMMYMMYMMYMMYM 5′ MMMXMMMXMMMXMMMXMMM 3′YMMMYMMMYMMMYMMMYMM 5′ XMMMXMMMXMMMXMMMXMM 3′ MMMYMMMYMMMYMMMYMMM

Nucleic acid molecules with a blunt 5′-end and 3′-end overhang end witha single modified nucleic acid.

Nucleic acid molecules with a 5′-end overhang and a blunt 3′-end with asingle modified nucleic acid.

Nucleic acid molecules with overhangs on both ends and all overhangs aremodified nucleic acids. In the pattern immediately below, M is n numberof modified nucleic acids, where n is an integer from 0 to 8 (i.e., 0,1, 2, 3, 4, 5, 6, 7 and 8).

5′  XXXXXXXXXXXXXXXXXXXM 3′ MYYYYYYYYYYYYYYYYYYY

Nucleic acid molecules with overhangs on both ends and some overhangnucleotides are modified nucleotides. In the patterns immediately below,M is n number of modified nucleotides, x is n number of unmodifiedoverhang nucleotides in the sense strand, y is n number of unmodifiedoverhang nucleotides in the antisense strand, where each n isindependently an integer from 0 to 8 (i.e., 0, 1, 2, 3, 4, 5, 6, 7 and8), and where each overhang is maximum of 20 nucleotides; preferably amaximum of 8 nucleotides (modified and/or unmodified).

5′          XXXXXXXXXXXXXXXXXXXM 3′         yYYYYYYYYYYYYYYYYYYY 5′         XXXXXXXXXXXXXXXXXXXMx 3′         yYYYYYYYYYYYYYYYYYYY 5′         XXXXXXXXXXXXXXXXXXXMxM 3′         yYYYYYYYYYYYYYYYYYYY 5′         XXXXXXXXXXXXXXXXXXXMxMx 3′         yYYYYYYYYYYYYYYYYYYY 5′         XXXXXXXXXXXXXXXXXXXMxMxM 3′         yYYYYYYYYYYYYYYYYYYY 5′         XXXXXXXXXXXXXXXXXXXMxMxMx 3′         yYYYYYYYYYYYYYYYYYYY 5′         XXXXXXXXXXXXXXXXXXXMxMxMxM 3′         yYYYYYYYYYYYYYYYYYYY 5′         XXXXXXXXXXXXXXXXXXXMxMxMxMx 3′         yYYYYYYYYYYYYYYYYYYY 5′        MXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYy 5′       xMXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYy 5′      MxMXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYy 5′     xMxMXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYy 5′    MxMxMXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYy 5′   xMxMxMXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYy 5′  MxMxMxMXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYy 5′ xMxMxMxMXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYy 5′        xXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYM 5′        xXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYMy 5′        xXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYMyM 5′        xXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYMyMy 5′        xXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYMyMyM 5′        xXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYMyMyMy 5′        xXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYMyMyMyM 5′        xXXXXXXXXXXXXXXXXXXX 3′          YYYYYYYYYYYYYYYYYYYMyMyMyMy 5′        XXXXXXXXXXXXXXXXXXXx 3′        MYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXXx 3′       yMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXXx 3′      MyMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXXx 3′     yMyMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXXx 3′    MyMyMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXXx 3′   yMyMyMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXXx 3′  MyMyMyMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXXx 3′ yMyMyMyMYYYYYYYYYYYYYYYYYYY

Modified nucleotides at the 3′ end of the sense region.

5′ XXXXXXXXXXXXXXXXXXXM 3′ YYYYYYYYYYYYYYYYYYY 5′ XXXXXXXXXXXXXXXXXXXMM3′ YYYYYYYYYYYYYYYYYYY 5′ XXXXXXXXXXXXXXXXXXXMMM 3′ YYYYYYYYYYYYYYYYYYY5′ XXXXXXXXXXXXXXXXXXXMMMM 3′ YYYYYYYYYYYYYYYYYYY 5′XXXXXXXXXXXXXXXXXXXMMMMM 3′ YYYYYYYYYYYYYYYYYYY 5′XXXXXXXXXXXXXXXXXXXMMMMMM 3′ YYYYYYYYYYYYYYYYYYY 5′XXXXXXXXXXXXXXXXXXXMMMMMMMM 3′ YYYYYYYYYYYYYYYYYYY 5′XXXXXXXXXXXXXXXXXXXMMMMMMMM 3′ YYYYYYYYYYYYYYYYYYY

Overhang at the 5′ end of the sense region.

5′        MXXXXXXXXXXXXXXXXXXX 3′         YYYYYYYYYYYYYYYYYYY 5′      MMXXXXXXXXXXXXXXXXXXX 3′         YYYYYYYYYYYYYYYYYYY 5′     MMMXXXXXXXXXXXXXXXXXXX 3′         YYYYYYYYYYYYYYYYYYY 5′    MMMMXXXXXXXXXXXXXXXXXXX 3′         YYYYYYYYYYYYYYYYYYY 5′   MMMMMXXXXXXXXXXXXXXXXXXX 3′         YYYYYYYYYYYYYYYYYYY 5′  MMMMMMXXXXXXXXXXXXXXXXXXX 3′         YYYYYYYYYYYYYYYYYYY 5′ MMMMMMMXXXXXXXXXXXXXXXXXXX 3′         YYYYYYYYYYYYYYYYYYY 5′MMMMMMMMXXXXXXXXXXXXXXXXXXX 3′         YYYYYYYYYYYYYYYYYYY

Overhang at the 3′ end of the antisense region.

5′         XXXXXXXXXXXXXXXXXXX 3′        MYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXX 3′       MMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXX 3′      MMMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXX 3′     MMMMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXX 3′    MMMMMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXX 3′   MMMMMMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXX 3′  MMMMMMMYYYYYYYYYYYYYYYYYYY 5′        XXXXXXXXXXXXXXXXXXX 3′ MMMMMMMMYYYYYYYYYYYYYYYYYYY

Modified nucleotide(s) within the sense region

5′   XXXXXXXXXMXXXXXXXXX 3′   YYYYYYYYYYYYYYYYYYY 5′  XXXXXXXXXXXXXXXXXXX 3′   YYYYYYYYYMYYYYYYYYY 5′  XXXXXXXXXXXXXXXXXXXMM 3′   YYYYYYYYYYYYYYYYYYY 5′  XXXXXXXXXXXXXXXXXXX 3′ MMYYYYYYYYYYYYYYYYYYY

Exemplary nucleic acid molecules are provided below along with theequivalent general structure in line with the symbols used above:

siHSP47-C siRNA to human and rat hsp47 having a 19 nucleotide (i.e.,19mer) duplex region and modified 2 nucleotide (i.e., deoxynucleotide)overhangs at the 3′-ends of the sense and antisense strands.

5′     GGACAGGCCUCUACAACUAdTdT 3′ 3′ dTdTCCUGUCCGGAGAUGUUGAU 5′ 5′  XXXXXXXXXXXXXXXXXXXMM 3′ MMYYYYYYYYYYYYYYYYYYY

siHSP47-Cd siRNA to human and rat hsp47 having a 25-mer duplex region, a2 nucleotide overhang at the 3′-end of the antisense strand and 2modified nucleotides at the 5′-terminal and penultimate positions of thesense strand.

5′   GGACAGGCCUCUACAACUACUACdGdA 3′ 3′ UUCCUGUCCGGAGAUGUUGAUGAUGCU 5′ 5′  XXXXXXXXXXXXXXXXXXXXXXXMM 3′ 3′ yyYYYYYYYYYYYYYYYYYYYYYYYYY 5′

siHSP47-1 siRNA to human and rat hsp47 cDNA 719-737 having a 19-merduplex region, and modified 2 nucleotide (i.e., deoxynucleotide)overhangs at the 3′-ends of the sense and antisense strands.

5′     CAGGCCUCUACAACUACUAdTdT 3′ 3′ dTdTGUCCGGAGAUGUUGAUGAU 5′ 5′    XXXXXXXXXXXXXXXXXXXMM 3′ 3′   MMYYYYYYYYYYYYYYYYYYY 5′

siHSP47-1d siRNA to human hsp47 cDNA 719-743 having a 25-mer with ablunt end at the 3′-end of the sense strand and a 2 nucleotide overhangat the 3′-end of the antisense strand, and 2 modified nucleotides at the5′-terminal and penultimate positions of the sense strand.

5′   CAGGCCUCUACAACUACUACGACdGdA 3′ 3′ UUGUCCGGAGAUGUUGAUGAUGCUGCU 5′ 5′  XXXXXXXXXXXXXXXXXXXXXXXMM 3′ 3′ yyYYYYYYYYYYYYYYYYYYYYYYYYY 5′

siHSP47-2 siRNA to human hsp47 cDNA 469-487 having a 19-mer duplexregion, and modified 2 nucleotide (i.e., deoxynucleotide) overhangs atthe 3′-ends of the sense and antisense strands.

5′     GAGCACUCCAAGAUCAACUdTdT 3′ 3′ dTdTCUCGUGAGGUUCUAGUUGA 5′ 5′    XXXXXXXXXXXXXXXXXXXMM 3′ 3′   MMYYYYYYYYYYYYYYYYYYY 5′

siHSP47-2d siRNA to human hsp47 cDNA 469-493 having a 25-mer duplexregion with a blunt end at the 3′-end of the sense strand and a 2nucleotide overhang at the 3′-end of the antisense strand, and 2modified nucleotides at the 5′-terminal and penultimate positions of thesense strand.

5′   GAGCACUCCAAGAUCAACUUCCGdCdG 3′ 3′ UUCUCGUGAGGUUCUAGUUGAAGGCGC 5′ 5′  XXXXXXXXXXXXXXXXXXXXXXXMM 3′ 3′ yyYYYYYYYYYYYYYYYYYYYYYYYYY 5′

siHSP47-2d rat siRNA to rat Gp46 cDNA 466-490 having a 25-mer duplexregion with a blunt end at the 3′-end of the sense strand and a 2nucleotide overhang at the 3′-end of the antisense strand, and 2modified nucleotides at the 5′-terminal and penultimate positions of thesense strand.

5′   GAACACUCCAAGAUCAACUUCCGdAdG 3′ 3′ UUCUUGUGAGGUUCUAGUUGAAGGCUC 5′ 5′  XXXXXXXXXXXXXXXXXXXXXXXMM 3′ 3′ yyYYYYYYYYYYYYYYYYYYYYYYYYY 5′

siHSP47-3 siRNA to human hsp47 cDNA 980-998 having a 19-mer duplexregion, and modified 2 nucleotide (i.e., deoxynucleotide) overhangs atthe 3′-ends of the sense and antisense strands.

5′     CTGAGGCCATTGACAAGAAdTdT 3′ 3′ dTdTGACUCCGGUAACUGUUCUU 5′ 5′    XXXXXXXXXXXXXXXXXXXMM 3′ 3′   MMYYYYYYYYYYYYYYYYYYY 5′

siHSP47-3d siRNA to human hsp47 cDNA 980-1004 having a 25-mer duplexregion with a blunt end at the 3′-end of the sense strand and a 2nucleotide overhang at the 3′-end of the antisense strand, and 2modified nucleotides at the 5′-terminal and penultimate positions of thesense strand.

5′   CTGAGGCCATTGACAAGAACAAGdGdC 3′ 3′ UUGACUCCGGUAACUGUUCUUGUUCCG 5′ 5′  XXXXXXXXXXXXXXXXXXXXXXXMM 3′ 3′ yyYYYYYYYYYYYYYYYYYYYYYYYYY 5′

siHSP47-4 siRNA to human hsp47 cDNA 735-753 having a 19-mer duplexregion, and modified 2 nucleotide (i.e., deoxynucleotide) overhangs atthe 3′-ends of the sense and antisense strands.

5′     CUACGACGACGAGAAGGAAdTdT 3′ 3′ dTdTGAUGCUGCUGCUCUUCCUU 5′ 5′    XXXXXXXXXXXXXXXXXXXMM 3′ 3′   MMYYYYYYYYYYYYYYYYYYY 5′

siHSP47-4-d siRNA to human hsp47 cDNA 735-759 having a 25-mer duplexregion with a blunt end at the 3′-end of the sense strand and a 2nucleotide overhang at the 3′-end of the antisense strand, and 2modified nucleotides at the 5′-terminal and penultimate positions of thesense strand.

5′   CUACGACGACGAGAAGGAAAAGCdTdG 3′ 3′ UUGAUGCUGCUGCUCUUCCUUUUCGAC 5′ 5′  XXXXXXXXXXXXXXXXXXXXXXXMM 3′ 3′ yyYYYYYYYYYYYYYYYYYYYYYYYYY 5′

siHSP47-5 siRNA to human hsp47 cDNA 621-639 having a 19-mer duplexregion, and modified 2 nucleotide (i.e., deoxynucleotide) overhangs atthe 3′-ends of the sense and antisense strands.

5′     GCCACACUGGGAUGAGAAAdTdT 3′ 3′ dTdTCGGUGUGACCCUACUCUUU 5′ 5′    XXXXXXXXXXXXXXXXXXXMM 3′ 3′   MMYYYYYYYYYYYYYYYYYYY 5′

siHSP47-6 siRNA to human hsp47 cDNA 446-464 having a 19-mer duplexregion, and modified 2 nucleotide (i.e., deoxynucleotide) overhangs atthe 3′-ends of the sense and antisense strands.

5′     GCAGCAAGCAGCACUACAAdTdT 3′ 3′ dTdTCGUCGUUCGUCGUGAUGUU 5′ 5′    XXXXXXXXXXXXXXXXXXXMM 3′ 3′   MMYYYYYYYYYYYYYYYYYYY 5′

siHSP47-7 siRNA to human hsp47 cDNA 692-710 having a 19-mer duplexregion, and modified 2 nucleotide (i.e., deoxynucleotide) overhangs atthe 3′-ends of the sense and antisense strands.

5′     CCGUGGGUGUCAUGAUGAUdTdT 3′ 3′ dTdTGGCACCCACAGUACUACUA 5′ 5′    XXXXXXXXXXXXXXXXXXXMM 3′ 3′   MMYYYYYYYYYYYYYYYYYYY 5′

Nicks and Gaps in Nucleic Acid Strands

Nucleic acid molecules (e.g., siNA molecules) provided herein may have astrand, preferably the sense strand, that is nicked or gapped. As such,nucleic acid molecules may have three or more strand, for example, suchas a meroduplex RNA (mdRNA) disclosed in International PatentApplication No. PCT/US07/081,836. Nucleic acid molecules with a nickedor gapped strand may be between about 1-49 nucleotides, or may be RISClength (e.g., about 15 to 25 nucleotides) or Dicer substrate length(e.g., about 25 to 30 nucleotides) such as disclosed herein.

Nucleic acid molecules with three or more strands include, for example,an ‘A’ (antisense) strand, ‘S1’ (second) strand, and ‘S2’ (third) strandin which the ‘S1’ and ‘S2’ strands arc complementary to and form basepairs with non-overlapping regions of the ‘A’ strand (e.g., an mdRNA canhave the form of A:S1S2). The S1, S2, or more strands together form whatis substantially similar to a sense strand to the ‘A’ antisense strand.The double-stranded region formed by the annealing of the ‘S1’ and ‘A’strands is distinct from and non-overlapping with the double-strandedregion formed by the annealing of the ‘S2’ and ‘A’ strands. An nucleicacid molecule (e.g., an siNA molecule) may be a “gapped” molecule,meaning a “gap” ranging from 0 nucleotides up to about 10 nucleotides(e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 nucleotides). Preferably, thesense strand is gapped. In some embodiments, the A:S1 duplex isseparated from the A:S2 duplex by a gap resulting from at least oneunpaired nucleotide (up to about 10 unpaired nucleotides) in the ‘A’strand that is positioned between the A:S I duplex and the A:S2 duplexand that is distinct from any one or more unpaired nucleotide at the3′-end of one or more of the ‘A’, ‘S1’, or ‘S2 strands. The A:S1 duplexmay be separated from the A: B2 duplex by a gap of zero nucleotides(i.e., a nick in which only a phosphodiester bond between twonucleotides is broken or missing in the polynucleotide molecule) betweenthe A:S1 duplex and the A:S2 duplex-which can also be referred to asnicked dsRNA (ndsRNA). For example, A: S1 S2 may be include a dsRNAhaving at least two double-stranded regions that combined total about 14base pairs to about 40 base pairs and the double-stranded regions areseparated by a gap of about 0 to about 10 nucleotides, optionally havingblunt ends, or A:S1S2 may include a dsRNA having at least twodouble-stranded regions separated by a gap of up to 10 nucleotideswherein at least one of the double-stranded regions includes betweenabout 5 base pairs and 13 base pairs.

Dicer Substrates

In certain embodiments, the nucleic acid molecules (e.g., siNAmolecules) provided herein may be a precursor “Dicer substrate”molecule, e.g., double stranded nucleic acid, processed in vivo toproduce an active nucleic acid molecules, for example as described inRossi, US Patent App. No. 20050244858. In certain conditions andsituations, it has been found that these relatively longer dsRNA siNAspecies, e.g., of from about 25 to about 30 nucleotides, can giveunexpectedly effective results in terms of potency and duration ofaction. Without wishing to be bound by any particular theory, it isthought that the longer dsRNA species serve as a substrate for theenzyme Dicer in the cytoplasm of a cell. In addition to cleaving doublestranded nucleic acid into shorter segments, Dicer may facilitate theincorporation of a single-stranded cleavage product derived from thecleaved dsRNA into the RNA-induced silencing complex (RISC complex) thatis responsible for the destruction of the cytoplasmic RNA derived fromthe target gene.

Dicer substrates may have certain properties which enhance itsprocessing by Dicer. Dicer substrates are of a length sufficient suchthat it is processed by Dicer to produce an active nucleic acid moleculeand may further include one or more of the following properties: (i) thedsRNA is asymmetric, e.g., has a 3′ overhang on the first strand(antisense strand) and (ii) the dsRNA has a modified 3′ end on theantisense strand (sense strand) to direct orientation of Dicer bindingand processing of the dsRNA to an active siRNA. In certain embodiments,the longest strand in the Dicer substrate may be 24-30 nucleotides.

Dicer substrates may be symmetric or asymmetric. The Dicer substrate mayhave a sense strand includes 22-28 nucleotides and the antisense strandmay include 24-30 nucleotides; thus, in some embodiments the resultingDicer substrate may have an overhang on the 3′ end of the antisensestrand. Dicer substrate may have a sense strand 25 nucleotides inlength, and the antisense strand having 27 nucleotides in length with a2 base 3′-overhang. The overhang may be 1-3 nucleotides, for example 2nucleotides. The sense strand may also have a 5′ phosphate.

An asymmetric Dicer substrate may further contain two deoxynucleotidesat the 3′-end of the sense strand in place of two of theribonucleotides. Some exemplary Dicer substrates lengths and structuresare 21+0, 21+2, 21−2, 22+0, 22+1, 22−1, 23+0, 23+2, 23−2, 24+0, 24+2,24−2, 25+0, 25+2, 25−2, 26+0, 26+2, 26−2, 27+0, 27+2, and 27−2.

The sense strand of a Dicer substrate may be between about 22 to about30 (e.g., about 22, 23, 24, 25, 26, 27, 28, 29 or 30); about 22 to about28; about 24 to about 30; about 25 to about 30; about 26 to about 30;about 26 and 29; or about 27 to about 28 nucleotides in length. Incertain preferred embodiments Dicer substrates contain sense andantisense strands, that are at least about 25 nucleotides in length andno longer than about 30 nucleotides; between about 26 and 29nucleotides; or 27 nucleotides in length. The sense and antisensestrands may be the same length (blunt ended), different lengths (haveoverhangs), or a combination. The sense and antisense strands may existon the same polynucleotide or on different polynucleotides. A Dicersubstrate may have a duplex region of about 19, 20, 21, 22, 23, 24, 25or 27 nucleotides.

Like other siNA molecules provided herein, the antisense strand of aDicer substrate may have any sequence that anneals to the antisensestrand under biological conditions, such as within the cytoplasm of aeukaryotic cell.

Dicer substrates may have any modifications to the nucleotide base,sugar or phosphate backbone as known in the art and/or as describedherein for other nucleic acid molecules (such as siNA molecules). Incertain embodiments, Dicer substrates may have a sense strand ismodified for Dicer processing by suitable modifiers located at the 3′end of the sense strand, i.e., the dsRNA is designed to directorientation of Dicer binding and processing. Suitable modifiers includenucleotides such as deoxyribonucleotides, dideoxyribonucleotides,acyclonucleotides and the like and sterically hindered molecules, suchas fluorescent molecules and the like. Acyclonucleotides substitute a2-hydroxyethoxymethyl group for-the 2′-deoxyribofuranosyl sugar normallypresent in dNMPs. Other nucleotides modifiers that could be used inDicer substrate siNA molecules include 3′-deoxyadenosine (cordycepin),3′-azido-3′-deoxythymidine (AZT), 2′,3′-dideoxyinosine (ddI),2′,3′-dideoxy-3′-thiacytidine (3TC),2′,3′-didehydro-2′,3′-dideoxythymidine (d4T) and the monophosphatenucleotides of 3′-azido-3′-deoxythymidine (AZT),2′,3′-dideoxy-3′-thiacytidine (3TC) and2′,3′-didehydro-2′,3′-dide-oxythymidine (d4T). In one embodiment,deoxynucleotides are used as the modifiers. When nucleotide modifiersare utilized, they may replace ribonucleotides (e.g., 1-3 nucleotidemodifiers, or 2 nucleotide modifiers are substituted for theribonucleotides on the 3′ end of the sense strand) such that the lengthof the Dicer substrate does not change. When sterically hinderedmolecules are utilized, they may be attached to the ribonucleotide atthe 3′ end of the antisense strand. Thus, in certain embodiments thelength of the strand does not change with the incorporation of themodifiers. In certain embodiments, two DNA bases in the dsRNA aresubstituted to direct the orientation of Dicer processing of theantisense strand. In a further embodiment of two terminal DNA bases aresubstituted for two ribonucleotides on the 3′-end of the sense strandforming a blunt end of the duplex on the 3′ end of the sense strand andthe 5′ end of the antisense strand, and a two-nucleotide RNA overhang islocated on the 3′-end of the antisense strand. This is an asymmetriccomposition with DNA on the blunt end and RNA bases on the overhangingend.

In certain embodiments modifications are included in the Dicer substratesuch that the modification does not prevent the nucleic acid moleculefrom serving as a substrate for Dicer. In one embodiment, one or moremodifications are made that enhance Dicer processing of the Dicersubstrate. One or more modifications may be made that result in moreeffective RNAi generation. One or more modifications may be made thatsupport a greater RNAi effect. One or more modifications are made thatresult in greater potency per each Dicer substrate to be delivered tothe cell. Modifications may be incorporated in the 3′-terminal region,the 5′-terminal region, in both the 3′-terminal and 5′-terminal regionor at various positions within the sequence. Any number and combinationof modifications can be incorporated into the Dicer substrate so long asthe modification does not prevent the nucleic acid molecule from servingas a substrate for Dicer. Where multiple modifications are present, theymay be the same or different. Modifications to bases, sugar moieties,the phosphate backbone, and their combinations are contemplated. Either5′-terminus can be phosphorylated.

Examples of Dicer substrate phosphate backbone modifications includephosphonates, including methylphosphonate, phosphorothioate, andphosphotriester modifications such as alkylphosphotriesters, and thelike. Examples of Dicer substrate sugar moiety modifications include2′-alkyl pyrimidine, such as 2′-O-methyl, 2′-fluoro, amino, and deoxymodifications and the like (see, e.g., Amarzguioui et al., 2003).Examples of Dicer substrate base group modifications include abasicsugars, 2-O-alkyl modified pyrimidines, 4-thiouracil, 5-bromouracil,5-iodouracil, and 5-(3-aminoallyl)-uracil and the like. Locked nucleicacids, or LNA's, could also be incorporated.

The sense strand may be modified for Dicer processing by suitablemodifiers located at the 3′ end of the sense strand, i.e., the Dicersubstrate is designed to direct orientation of Dicer binding andprocessing. Suitable modifiers include nucleotides such asdeoxyribonucleotides, dideoxyribonucleotides, acyclonucleotides and thelike and sterically hindered molecules, such as fluorescent moleculesand the like. Acyclonucleotides substitute a 2-hydroxyethoxymethyl groupfor-the 2′-deoxyribofuranosyl sugar normally present in dNMPs. Othernucleotides modifiers could include 3′-deoxyadenosine (cordycepin),3′-azido-3′-deoxythymidine (AZT), 2′,3′-dideoxyinosine (ddI),2′,3′-dideoxy-3′-thiacytidine (3TC),2′,3′-didehydro-2′,3′-dideoxythymidine (d4T) and the monophosphatenucleotides of 3′-azido-3′-deoxythymidine (AZT),2′,3′-dideoxy-3′-thiacytidine (3TC) and2′,3′-didehydro-2′,3′-dide-oxythymidine (d4T). In one embodiment,deoxynucleotides are used as the modifiers. When nucleotide modifiersare utilized, 1-3 nucleotide modifiers, or 2 nucleotide modifiers aresubstituted for the ribonucleotides on the 3′ end of the sense strand.When sterically hindered molecules are utilized, they are attached tothe ribonucleotide at the 3′ end of the antisense strand. Thus, thelength of the strand does not change with the incorporation of themodifiers. In another embodiment, the invention contemplatessubstituting two DNA bases in the Dicer substrate to direct theorientation of Dicer processing of the antisense strand. In a furtherembodiment of the present invention, two terminal DNA bases aresubstituted for two ribonucleotides on the 3′-end of the sense strandforming a blunt end of the duplex on the 3′ end of the sense strand andthe 5′ end of the antisense strand, and a two-nucleotide RNA overhang islocated on the 3′-end of the antisense strand. This is an asymmetriccomposition with DNA on the blunt end and RNA bases on the overhangingend.

The antisense strand may be modified for Dicer processing by suitablemodifiers located at the 3′ end of the antisense strand, i.e., the dsRNAis designed to direct orientation of Dicer binding and processing.Suitable modifiers include nucleotides such as deoxyribonucleotides,dideoxyribonucleotides, acyclonucleotides and the like and stericallyhindered molecules, such as fluorescent molecules and the like.Acyclonucleotides substitute a 2-hydroxyethoxymethyl group for the2′-deoxyribofuranosyl sugar normally present in dNMPs. Other nucleotidemodifiers could include 3′-deoxyadenosine (cordycepin),3′-azido-3′-deoxythymidine (AZT), 2′,3′-dideoxyinosine (ddI),2′,3′-dideoxy-3′-thiacytidine (3TC),2′,3′-didehydro-2′,3′-dideoxythymidine (d4T) and the monophosphatenucleotides of 3′-azido-3′-deoxythymidine (AZT),2′,3′-dideoxy-3′-thiacytidine (3TC) and2′,3′-didehydro-2′,3′-dide-oxythymidine (d4T). In one embodiment,deoxynucleotides are used as the modifiers. When nucleotide modifiersare utilized, 1-3 nucleotide modifiers, or 2 nucleotide modifiers aresubstituted for the ribonucleotides on the 3′ end of the antisensestrand. When sterically hindered molecules are utilized, they areattached to the ribonucleotide at the 3′ end of the antisense strand.Thus, the length of the strand does not change with the incorporation ofthe modifiers. In another embodiment, the invention contemplatessubstituting two DNA bases in the dsRNA to direct the orientation ofDicer processing. In a further invention, two terminal DNA bases arelocated on the 3′ end of the antisense strand in place of tworibonucleotides forming a blunt end of the duplex on the 5′ end of thesense strand and the 3′ end of the antisense strand, and atwo-nucleotide RNA overhang is located on the 3′-end of the sensestrand. This is an asymmetric composition with DNA on the blunt end andRNA bases on the overhanging end.

Dicer substrates with a sense and an antisense strand can be linked by athird structure. The third structure will not block Dicer activity onthe Dicer substrate and will not interfere with the directed destructionof the RNA transcribed from the target gene. The third structure may bea chemical linking group. Suitable chemical linking groups are known inthe art and can be used. Alternatively, the third structure may be anoligonucleotide that links the two oligonucleotides of the dsRNA is amanner such that a hairpin structure is produced upon annealing of thetwo oligonucleotides making up the Dicer substrate. The hairpinstructure preferably does not block Dicer activity on the Dicersubstrate or interfere with the directed destruction of the RNAtranscribed from the target gene.

The sense and antisense strands of the Dicer substrate are not requiredto be completely complementary. They only need to be substantiallycomplementary to anneal under biological conditions and to provide asubstrate for Dicer that produces an siRNA sufficiently complementary tothe target sequence.

Dicer substrate can have certain properties that enhance its processingby Dicer. The Dicer substrate can have a length sufficient such that itis processed by Dicer to produce an active nucleic acid molecules (e.g.,siRNA) and may have one or more of the following properties: (i) theDicer substrate is asymmetric, e.g., has a 3′ overhang on the firststrand (antisense strand) and (ii) the Dicer substrate has a modified 3′end on the second strand (sense strand) to direct orientation of Dicerbinding and processing of the Dicer substrate to an active siRNA. TheDicer substrate can be asymmetric such that the sense strand includes22-28 nucleotides and the antisense strand includes 24-30 nucleotides.Thus, the resulting Dicer substrate has an overhang on the 3′ end of theantisense strand. The overhang is 1-3 nucleotides, for example 2nucleotides. The sense strand may also have a 5′ phosphate.

A Dicer substrate may have an overhang on the 3′ end of the antisensestrand and the sense strand is modified for Dicer processing. The 5′ endof the sense strand may have a phosphate. The sense and antisensestrands may anneal under biological conditions, such as the conditionsfound in the cytoplasm of a cell. A region of one of the strands,particularly the antisense strand, of the Dicer substrate may have asequence length of at least 19 nucleotides, wherein these nucleotidesare in the 21-nucleotide region adjacent to the 3′ end of the antisensestrand and are sufficiently complementary to a nucleotide sequence ofthe RNA produced from the target gene. A Dicer substrate may also haveone or more of the following additional properties: (a) the antisensestrand has a right shift from a corresponding 21-mer (i.e., theantisense strand includes nucleotides on the right side of the moleculewhen compared to the corresponding 21-mer), (b) the strands may not becompletely complementary, i.e., the strands may contain simple mismatchpairings and (c) base modifications such as locked nucleic acid(s) maybe included in the 5′ end of the sense strand.

An antisense strand of a Dicer substrate nucleic acid molecule may bemodified to include 1-9 ribonucleotides on the 5′-end to give a lengthof 22-28 nucleotides. When the antisense strand has a length of 21nucleotides, then 1-7 ribonucleotides, or 2-5 ribonucleotides and or 4ribonucleotides may be added on the 3′-end. The added ribonucleotidesmay have any sequence. Although the added ribonucleotides may becomplementary to the target gene sequence, full complementarity betweenthe target sequence and the antisense strands is not required. That is,the resultant antisense strand is sufficiently complementary with thetarget sequence. A sense strand may then have 24-30 nucleotides. Thesense strand may be substantially complementary with the antisensestrand to anneal to the antisense strand under biological conditions. Inone embodiment, the antisense strand may be synthesized to contain amodified 3′-end to direct Dicer processing. The sense strand may have a3′ overhang. The antisense strand may be synthesized to contain amodified 3′-end for Dicer binding and processing and the sense strandhas a 3′ overhang.

Heat Shock Protein 47

Heat shock protein 47 (HSP47) is a collagen-specific molecular chaperoneand resides in the endoplasmic reticulum. It interacts with procollagenduring the process of folding, assembling and transporting from theendoplasmic reticulum (Nagata Trends Biochem Sci 1996; 21:22-6; Razzaqueet al. 2005; Contrib Nephrol 2005; 148: 57-69; Koide et al. 2006 J.Biol. Chem.; 281: 3432-38; Leivo et al. Dev. Biol. 1980; 76:100-114;Masuda et al. J. Clin. Invest. 1994; 94:2481-2488; Masuda et al. CellStress Chaperones 1998; 3:256-264). HSP47 has been reported to have anupregulated expression in various tissue fibrosis (Koide et al. J BiolChem 1999; 274: 34523-26), such as liver cirrhosis (Masuda et al. J ClinInvest 1994; 94:2481-8), pulmonary fibrosis (Razzaque et al. VirchowsArch 1998; 432:455-60; Kakugawa ct al. Eur Respir J 2004; 24: 57-65),and glomerulosclerosis (Moriyama et al. Kidney Int 1998; 54: 110-19).Exemplary nucleic acid sequence of target human hsp47 cDNA is disclosedin GenBank accession number: NM_(—)001235 and the corresponding mRNAsequence, for example as listed as SEQ ID NO: 1. One of ordinary skillin the art would understand that a given sequence may change over timeand to incorporate any changes needed in the nucleic acid moleculesherein accordingly.

The specific association of HSP47 with a diverse range of collagen typesmakes HSP47 a potential target for the treatment of fibrosis. Inhibitionof hsp47 expression may prevent extracellular collagen I secretion. Satoet al. (Nat Biotechnol 2008; 26:431-442) explored this possibility byusing siRNA for the inhibition hsp47 expression and preventing theprogression of hepatic fibrosis in rats. Similarly, Chen et al. (Br JDermatol 2007; 156: 1188-1195) and Wang et al. (Plast. Reconstr Surg2003; 111: 1980-7) investigated the inhibition hsp47 expression by RNAinterference technology.

Methods and Compositions for Inhibiting hsp47

Provided are compositions and methods for inhibition of hsp47 expressionby using small nucleic acid molecules, such as short interfering nucleicacid (siNA), interfering RNA (RNAi), short interfering RNA (siRNA),double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA(shRNA) molecules capable of mediating or that mediate RNA interferenceagainst hsp47 gene expression. The composition and methods disclosedherein are also useful in treating various fibrosis such as liverfibrosis, lung fibrosis, and kidney fibrosis.

Nucleic acid molecule(s) and/or methods of the invention are used todown regulate the expression of gene(s) that encode RNA referred to, byexample, Genbank Accession NM_(—)001235.

Compositions, methods and kits provided herein may include one or morenucleic acid molecules (e.g., siNA) and methods that independently or incombination modulate (e.g., downregulate) the expression of hsp47protein and/or genes encoding hsp47 proteins, proteins and/or genesencoding hsp47 associated with the maintenance and/or development ofdiseases, conditions or disorders associated with hsp47, such as liverfibrosis, cirrhosis, pulmonary fibrosis, kidney fibrosis, peritonealfibrosis, chronic hepatic damage, and fibrillogenesis (e.g., genesencoding sequences comprising those sequences referred to by GenBankAccession Nos. NM_(—)001235), or a hsp47 gene family member where thegenes or gene family sequences share sequence homology. The descriptionof the various aspects and embodiments is provided with reference toexemplary gene hsp47. However, the various aspects and embodiments arealso directed to other related hsp47 genes, such as homolog genes andtranscript variants, and polymorphisms (e.g., single nucleotidepolymorphism, (SNPs)) associated with certain hsp47 genes. As such, thevarious aspects and embodiments are also directed to other genes thatare involved in hsp47 mediated pathways of signal transduction or geneexpression that are involved, for example, in the maintenance ordevelopment of diseases, traits, or conditions described herein. Theseadditional genes can be analyzed for target sites using the methodsdescribed for the hsp47 gene herein. Thus, the modulation of other genesand the effects of such modulation of the other genes can be performed,determined, and measured as described herein.

In one embodiment, compositions and methods provided herein include adouble-stranded short interfering nucleic acid (siNA) molecule thatdown-regulates expression of a hsp47 gene (e.g., human hsp47 exemplifiedby SEQ ID NO:1), where the nucleic acid molecule includes about 15 toabout 49 base pairs.

In one embodiment, a nucleic acid disclosed may be used to inhibit theexpression of the hsp47 gene or a hsp47 gene family where the genes orgene family sequences share sequence homology. Such homologous sequencescan be identified as is known in the art, for example using sequencealignments. Nucleic acid molecules can be designed to target suchhomologous sequences, for example using perfectly complementarysequences or by incorporating non-canonical base pairs, for examplemismatches and/or wobble base pairs, that can provide additional targetsequences. In instances where mismatches are identified, non-canonicalbase pairs (for example, mismatches and/or wobble bases) can be used togenerate nucleic acid molecules that target more than one gene sequence.In a non-limiting example, non-canonical base pairs such as UU and CCbase pairs are used to generate nucleic acid molecules that are capableof targeting sequences for differing hsp47 targets that share sequencehomology. As such, one advantage of using siNAs disclosed herein is thata single nucleic acid can be designed to include nucleic acid sequencethat is complementary to the nucleotide sequence that is conservedbetween the homologous genes. In this approach, a single nucleic acidcan be used to inhibit expression of more than one gene instead of usingmore than one nucleic acid molecule to target the different genes.

Nucleic acid molecules may be used to target conserved sequencescorresponding to a gene family or gene families such as hsp47 familygenes. As such, nucleic acid molecules targeting multiple hsp47 targetscan provide increased therapeutic effect. In addition, nucleic acid canbe used to characterize pathways of gene function in a variety ofapplications. For example, nucleic acid molecules can be used to inhibitthe activity of target gcnc(s) in a pathway to determine the function ofuncharacterized gene(s) in gene function analysis, mRNA functionanalysis, or translational analysis. The nucleic acid molecules can beused to determine potential target gene pathways involved in variousdiseases and conditions toward pharmaceutical development. The nucleicacid molecules can be used to understand pathways of gene expressioninvolved in, for example fibroses such as liver, kidney or pulmonaryfibrosis, and/or inflammatory and proliferative traits, diseases,disorders, and/or conditions.

In one embodiment, the compositions and methods provided herein includea nucleic acid molecule having RNAi activity against hsp47 RNA, wherethe nucleic acid molecule includes a sequence complementary to any RNAhaving hsp47 encoding sequence, such as those sequences having sequencesas shown in Table I. In another embodiment, a nucleic acid molecule mayhave RNAi activity against hsp47 RNA, where the nucleic acid moleculeincludes a sequence complementary to an RNA having variant hsp47encoding sequence, for example other mutant hsp47 genes not shown inTable I but known in the art to be associated with the maintenanceand/or development of fibrosis. Chemical modifications as shown in TableI or otherwise described herein can be applied to any nucleic acidconstruct disclosed herein. In another embodiment, a nucleic acidmolecule disclosed herein includes a nucleotide sequence that caninteract with nucleotide sequence of a hsp47 gene and thereby mediatesilencing of hsp47 gene expression, for example, wherein the nucleicacid molecule mediates regulation of hsp47 gene expression by cellularprocesses that modulate the chromatin structure or methylation patternsof the hsp47 gene and prevent transcription of the hsp47 gene.

Nucleic acid molecules disclosed herein may have RNAi activity againsthsp47 RNA, where the nucleic acid molecule includes a sequencecomplementary to any RNA having hsp47 encoding sequence, such as thosesequences having GenBank Accession Nos. NM_(—)001235. Nucleic acidmolecules may have RNAi activity against hsp47 RNA, where the nucleicacid molecule includes a sequence complementary to an RNA having varianthsp47 encoding sequence, for example other mutant hsp47 genes known inthe art to be associated with the maintenance and/or development offibrosis. Nucleic acid molecules disclosed herein include a nucleotidesequence that can interact with nucleotide sequence of a hsp47 gene andthereby mediate silencing of hsp47 gene expression, e.g., where thenucleic acid molecule mediates regulation of hsp47 gene expression bycellular processes that modulate the chromatin structure or methylationpatterns of the hsp47 gene and prevent transcription of the hsp47 gene.

Methods of Treatment

The specific association of HSP47 with a diverse range of collagen typesmakes hsp47 a target for the treatment of fibrosis. Inhibition of hsp47expression may prevent extracellular collagen I secretion. Sato et al.(Nat Biotechnol 2008; 26:431-442) explored this possibility by usingsiRNA for the inhibition hsp47 expression and preventing the progressionof hepatic fibrosis in rats. Similarly, Chen et al. (Br J Dermatol 2007;156: 1188-1195) and Wang et al. (Plast. Reconstr Surg 2003; 111: 1980-7)investigated the inhibition hsp47 expression by RNA interferencetechnology.

In one embodiment, nucleic acid molecules may be used to down regulateor inhibit the expression of hsp47 and/or hsp47 proteins arising fromhsp47 and/or hsp47 haplotype polymorphisms that are associated with adisease or condition, (e.g., fibrosis). Analysis of hsp47 and/or hsp47genes, or hsp47 and/or hsp47 protein or RNA levels can be used toidentify subjects with such polymorphisms or those subjects who are atrisk of developing traits, conditions, or diseases described herein.These subjects are amenable to treatment, for example, treatment withnucleic acid molecules disclosed herein and any other composition usefulin treating diseases related to hsp47 and/or hsp47 gene expression. Assuch, analysis of hsp47 and/or hsp47 protein or RNA levels can be usedto determine treatment type and the course of therapy in treating asubject. Monitoring of hsp47 and/or hsp47 protein or RNA levels can beused to predict treatment outcome and to determine the efficacy ofcompounds and compositions that modulate the level and/or activity ofcertain hsp47 and/or hsp47 proteins associated with a trait, condition,or disease.

Provided arc compositions and methods for inhibition of hsp47 expressionby using small nucleic acid molecules as provided herein, such as shortinterfering nucleic acid (siNA), interfering RNA (RNAi), shortinterfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA),and short hairpin RNA (shRNA) molecules capable of mediating or thatmediate RNA interference against hsp47 gene expression. The compositionand methods disclosed herein arc also useful in treating variousfibrosis such as liver fibrosis, lung fibrosis, and kidney fibrosis.

The nucleic acid molecules disclosed herein individually, or incombination or in conjunction with other drugs, can be use forpreventing or treating diseases, traits, conditions and/or disordersassociated with hsp47, such as liver fibrosis, cirrhosis, pulmonaryfibrosis, kidney fibrosis, peritoneal fibrosis, chronic hepatic damage,and fibrillogenesis.

The nucleic acid molecules disclosed herein are able to inhibit theexpression of hsp47 in a sequence specific manner. The nucleic acidmolecules may include a sense strand and an antisense strand whichinclude contiguous nucleotides that are at least partially complementary(antisense) to a hsp47 mRNA.

In some embodiments, dsRNA specific for hsp47 can be used in conjunctionwith other dsRNA specific for other molecular chaperones that assist inthe folding of newly synthesized proteins such as, calnexin,calreticulin, BiP (Bergeron et al. Trends Biochem. Sci. 1994;19:124-128; Herbert et al. 1995; Cold Spring Harb. Symp. Quant. Biol.60:405-415)

Fibrosis can be treated by RNA interference using nucleic acid moleculesas disclosed herein. Exemplary fibrosis include liver fibrosis,peritoneal fibrosis, lung fibrosis, kidney fibrosis. The nucleic acidmolecules disclosed herein may inhibit the expression of hsp47 in asequence specific manner.

Treatment of fibrosis can be monitored by determining the level ofextracellular collagen using suitable techniques known in the art suchas, using anti-collagen I antibodies. Treatment can also be monitored bydetermining the level of hsp47 mRNA or the level of HSP47 protein in thecells of the affected tissue. Treatment can also be monitored bynon-invasive scanning of the affected organ or tissue such as bycomputer assisted tomography scan, magnetic resonance elastographyscans.

A method for treating or preventing hsp47 associated disease orcondition in a subject or organism may include contacting the subject ororganism with a nucleic acid molecule as provided herein underconditions suitable to modulate the expression of the hsp47 gene in thesubject or organism.

A method for treating or preventing fibrosis in a subject or organismmay include contacting the subject or organism with a nucleic acidmolecule under conditions suitable to modulate the expression of thehsp47 gene in the subject or organism.

A method for treating or preventing one or more fibroses selected fromthe group consisting of liver fibrosis, kidney fibrosis, and pulmonaryfibrosis in a subject or organism may include contacting the subject ororganism with a nucleic acid molecule under conditions suitable tomodulate the expression of the hsp47 gene in the subject or organism.

Fibrotic Diseases

Fibrotic diseases are generally characterized by the excess depositionof a fibrous material within the extracellular matrix, which contributesto abnormal changes in tissue architecture and interferes with normalorgan function.

All tissues damaged by trauma respond by the initiation of awound-healing program. Fibrosis, a type of disorder characterized byexcessive scarring, occurs when the normal self-limiting process ofwound healing response is disturbed, and causes excessive production anddeposition of collagen. As a result, normal organ tissue is replacedwith scar tissue, which eventually leads to the functional failure ofthe organ.

Fibrosis may be initiated by diverse causes and in various organs. Livercirrhosis, pulmonary fibrosis, sarcoidosis, keloids and kidney fibrosisare all chronic conditions associated with progressive fibrosis, therebycausing a continuous loss of normal tissue function.

Acute fibrosis (usually with a sudden and severe onset and of shortduration) occurs as a common response to various forms of traumaincluding accidental injuries (particularly injuries to the spine andcentral nervous system), infections, surgery, ischemic illness (e.g.cardiac scarring following heart attack), burns, environmentalpollutants, alcohol and other types of toxins, acute respiratorydistress syndrome, radiation and chemotherapy treatments).

Fibrosis, a fibrosis related pathology or a pathology related toaberrant crosslinking of cellular proteins may all be treated by thesiRNAs disclosed herein. Fibrotic diseases or diseases in which fibrosisis evident (fibrosis related pathology) include both acute and chronicforms of fibrosis of organs, including all etiological variants of thefollowing: pulmonary fibrosis, including interstitial lung disease andfibrotic lung disease, liver fibrosis, cardiac fibrosis includingmyocardial fibrosis, kidney fibrosis including chronic renal failure,skin fibrosis including scleroderma, keloids and hypertrophic scars;myelofibrosis (bone marrow fibrosis); all types of ocular scarringincluding proliferative vitreoretinopathy (PVR) and scarring resultingfrom surgery to treat cataract or glaucoma; inflammatory bowel diseaseof variable etiology, macular degeneration, Grave's ophthalmopathy, druginduced ergotism, keloid scars, scleroderma, psoriasis, glioblastoma inLi-Fraumeni syndrome, sporadic glioblastoma, myleoid leukemia, acutemyelogenous leukemia, myelodysplastic syndrome, myeloproferativesyndrome, gynecological cancer, Kaposi's sarcoma, Hansen's disease, andcollagenous colitis.

In various embodiments, the compounds (nucleic acid molecules) asdisclosed herein may be used to treat fibrotic diseases, for example asdisclosed herein, as well as many other diseases and conditions apartfrom fibrotic diseases, for example such as disclosed herein. Otherconditions to be treated include fibrotic diseases in otherorgans—kidney fibrosis for any reason (CKD including ESRD); lungfibrosis (including ILF); myelofibrosis, abnormal scarring (keloids)associated with all possible types of skin injury accidental andjatrogenic (operations); scleroderma; cardiofibrosis, failure ofglaucoma filtering operation; intestinal adhesions.

Ocular Surgery and Fibrotic Complications

Contracture of scar tissue resulting from eye surgery may often occur.Glaucoma surgery to create new drainage channels often fails due toscarring and contraction of tissues and the generated drainage systemmay be blocked requiring additional surgical intervention. Currentanti-scarring regimens (Mitomycin C or 5FU) are limited due to thecomplications involved (e.g. blindness) e.g. see Cordeiro M F, et al.,Human anti-transforming growth factor-beta2 antibody: a new glaucomaanti-scarring agent Invest Ophthalmol V is Sci. 1999 September;40(10):2225-34. There may also be contraction of scar tissue formedafter corneal trauma or corneal surgery, for example laser or surgicaltreatment for myopia or refractive error in which contraction of tissuesmay lead to inaccurate results. Scar tissue may be formed on/in thevitreous humor or the retina, for example, and may eventually causesblindness in some diabetics, and may be formed after detachment surgery,called proliferative vitreoretinopathy (PVR). PVR is the most commoncomplication following retinal detachment and is associated with aretinal hole or break. PVR refers to the growth of cellular membraneswithin the vitreous cavity and on the front and back surfaces of theretina containing retinal pigment epithelial (RPE) cells. Thesemembranes, which are essentially scar tissues, exert traction on theretina and may result in recurrences of retinal detachment, even afteran initially successful retinal detachment procedure.

Scar tissue may be formed in the orbit or on eye and eyelid musclesafter squint, orbital or eyelid surgery, or thyroid eye disease, andwhere scarring of the conjunctiva occurs as may happen after glaucomasurgery or in cicatricial disease, inflammatory disease, for example,pemphigoid, or infective disease, for example, trachoma. A further eyeproblem associated with the contraction of collagen-including tissues isthe opacification and contracture of the lens capsule after cataractextraction. Important role for MMPs has been recognized in oculardiseases including wound healing, dry eye, sterile corneal ulceration,recurrent epithelial erosion, corneal neovascularization, pterygium,conjuctivochalasis, glaucoma, PVR, and ocular fibrosis.

Liver Fibrosis

Liver fibrosis (LF) is a generally irreversible consequence of hepaticdamage of several etiologies. In the Western world, the main etiologiccategories are: alcoholic liver disease (30-50%), viral hepatitis (30%),biliary disease (5-10%), primary hemochromatosis (5%), and drug-relatedand cryptogenic cirrhosis of, unknown etiology, (10-15%). Wilson'sdisease, α₁-antitrypsin deficiency and other rare diseases also haveliver fibrosis as one of the symptoms. Liver cirrhosis, the end stage ofliver fibrosis, frequently requires liver transplantation and is amongthe top ten causes of death in the Western world.

Kidney Fibrosis and Related Conditions.

Chronic Renal Failure (CRF)

Chronic renal failure is a gradual and progressive loss of the abilityof the kidneys to excrete wastes, concentrate urine, and conserveelectrolytes. CRF is slowly progressive. It most often results from anydisease that causes gradual loss of kidney function, and fibrosis is themain pathology that produces CRF.

Diabetic Nephropathy

Diabetic nephropathy, hallmarks of which are glomerulosclerosis andtubulointerstitial fibrosis, is the single most prevalent cause ofend-stage renal disease in the modern world, and diabetic patientsconstitute the largest population on dialysis. Such therapy is costlyand far from optimal. Transplantation offers a better outcome butsuffers from a severe shortage of donors.

Chronic Kidney Disease

Chronic kidney disease (CKD) is a worldwide public health problem and isrecognized as a common condition that is associated with an increasedrisk of cardiovascular disease and chronic renal failure (CRF).

The Kidney Disease Outcomes Quality Initiative (K/DOQI) of the NationalKidney Foundation (NKF) defines chronic kidney disease as either kidneydamage or a decreased kidney glomerular filtration rate (GFR) for threeor more months. Other markers of CKD are also known and used fordiagnosis. In general, the destruction of renal mass with irreversiblesclerosis and loss of nephrons leads to a progressive decline in GFR.Recently, the K/DOQI published a classification of the stages of CKD, asfollows:

Stage 1: Kidney damage with normal or increased GFR (>90 mL/min/1.73 m2)

Stage 2: Mild reduction in GFR (60-89 mL/min/1.73 m2)

Stage 3: Moderate reduction in GFR (30-59 mL/min./1.73 m2)

Stage 4: Severe reduction in GFR (15-29 ml./min/1.73 m2)

Stage 5: Kidney failure (GFR <15 mL/min/1.73 m2 or dialysis)

In stages 1 and 2 CKD, GFR alone does not confirm the diagnosis. Othermarkers of kidney damage, including abnormalities in the composition ofblood or urine or abnormalities in imaging tests, may be relied upon.

Pathophysiology of CKD

Approximately 1 million nephrons are present in each kidney, eachcontributing to the total GFR. Irrespective of the etiology of renalinjury, with progressive destruction of nephrons, the kidney is able tomaintain GFR by hyperfiltration and compensatory hypertrophy of theremaining healthy nephrons. This nephron adaptability allows forcontinued normal clearance of plasma solutes so that substances such asurea and creatinine start to show significant increases in plasma levelsonly after total GFR has decreased to 50%, when the renal reserve hasbeen exhausted. The plasma creatinine value will approximately doublewith a 50% reduction in GFR. Therefore, a doubling in plasma creatininefrom a baseline value of 0.6 mg/dL to 1.2 mg/dL in a patient actuallyrepresents a loss of 50% of functioning nephron mass.

The residual nephron hyperfiltration and hypertrophy, althoughbeneficial for the reasons noted, is thought to represent a major causeof progressive renal dysfunction. This is believed to occur because ofincreased glomerular capillary pressure, which damages the capillariesand leads initially to focal and segmental glomerulosclerosis andeventually to global glomerulosclerosis. This hypothesis has been basedon studies of five-sixths nephrectomized rats, which develop lesionsthat are identical to those observed in humans with CKD.

The two most common causes of chronic kidney disease are diabetes andhypertension. Other factors include acute insults from nephrotoxins,including contrasting agents, or decreased perfusion; Proteinuria;Increased renal ammoniagenesis with interstitial injury; Hyperlipidemia;Hyperphosphatemia with calcium phosphate deposition; Decreased levels ofnitrous oxide and smoking.

In the United States, the incidence and prevalence of CKD is rising,with poor outcomes and high cost to the health system. Kidney disease isthe ninth leading cause of death in the US. The high rate of mortalityhas led the US Surgeon General's mandate for America's citizenry,Healthy People 2010, to contain a chapter focused on CKD. The objectivesof this chapter are to articulate goals and to provide strategies toreduce the incidence, morbidity, mortality, and health costs of chronickidney disease in the United States.

The incidence rates of end-stage renal disease (ESRD) have alsoincreased steadily internationally since 1989. The United States has thehighest incident rate of ESRD, followed by Japan. Japan has the highestprevalence per million population, followed by the US.

The mortality rates associated with hemodialysis are striking andindicate that the life expectancy of patients entering into hemodialysisis markedly shortened. At every age, patients with ESRD on dialysis havesignificantly increased mortality when compared with nondialysispatients and individuals without kidney disease. At age 60 years, ahealthy person can expect to live for more than 20 years, whereas thelife expectancy of a 60-year-old patient starting hemodialysis is closerto 4 years (Aurora and Verelli, May 21, 2009. Chronic Renal Failure:Treatment & Medication. Emedicine.http://emedicine.medscape.com/article/238798-treatment).

Pulmonary Fibrosis

Interstitial pulmonary fibrosis (IPF) is scarring of the lung caused bya variety of inhaled agents including mineral particles, organic dusts,and oxidant gases, or by unknown reasons (idiopathic lung fibrosis). Thedisease afflicts millions of individuals worldwide, and there are noeffective therapeutic approaches. A major reason for the lack of usefultreatments is that few of the molecular mechanisms of disease have beendefined sufficiently to design appropriate targets for therapy (Lasky JA., Brody A R. (2000), “Interstitial fibrosis and growth factors”,Environ Health Perspect.;108 Suppl 4:751-62).

Cardiac Fibrosis

Heart failure is unique among the major cardiovascular disorders in thatit alone is increasing in prevalence while there has been a strikingdecrease in other conditions. Some of this can be attributed to theaging of the populations of the United States and Europe. The ability tosalvage patients with myocardial damage is also a major factor, as thesepatients may develop progression of left ventricular dysfunction due todeleterious remodelling of the heart.

The normal myocardium is composed of a variety of cells, cardiacmyocytes and noncardiomyocytes, which include endothelial and vascularsmooth muscle cells and fibroblasts.

Structural remodeling of the ventricular wall is a key determinant ofclinical outcome in heart disease. Such remodeling involves theproduction and destruction of extracellular matrix proteins, cellproliferation and migration, and apoptotic and necrotic cell death.Cardiac fibroblasts are crucially involved in these processes, producinggrowth factors and cytokines that act as autocrine and paracrinefactors, as well as extracellular matrix proteins and proteinases.Recent studies have shown that the interactions between cardiacfibroblasts and cardiomyocytes are essential for the progression ofcardiac remodeling of which the net effect is deterioration in cardiacfunction and the onset of heart failure (Manabe I, et al., (2002), “Geneexpression in fibroblasts and fibrosis: involvement in cardiachypertrophy”, Circ Res. 13; 91(12):1103-13).

Burns and Scars

A particular problem which may arise, particularly in fibrotic disease,is contraction of tissues, for example contraction of scars. Contractionof tissues including extracellular matrix components, especially ofcollagen-including tissues, may occur in connection with many differentpathological conditions and with surgical or cosmetic procedures.Contracture, for example, of scars, may cause physical problems, whichmay lead to the need for medical treatment, or it may cause problems ofa purely cosmetic nature. Collagen is the major component of scar andother contracted tissue and as such is the most important structuralcomponent to consider. Nevertheless, scar and other contracted tissuealso includes other structural components, especially otherextracellular matrix components, for example, elastin, which may alsocontribute to contraction of the tissue.

Contraction of collagen-including tissue, which may also include otherextracellular matrix components, frequently occurs in the healing ofburns. The burns may be chemical, thermal or radiation burns and may beof the eye, the surface of the skin or the skin and the underlyingtissues. It may also be the case that there are burns on internaltissues, for example, caused by radiation treatment. Contraction ofburnt tissues is often a problem and may lead to physical and/orcosmetic problems, for example, loss of movement and/or disfigurement.

Skin grafts may be applied for a variety of reasons and may oftenundergo contraction after application. As with the healing of burnttissues the contraction may lead to both physical and cosmetic problems.It is a particularly serious problem where many skin grafts are neededas, for example, in a serious burns case.

Contraction is also a problem in production of artificial skin. To makea true artificial skin it is necessary to have an epidermis made ofepithelial cells (keratinocytes) and a dermis made of collagen populatedwith fibroblasts. It is important to have both types of cells becausethey signal and stimulate each other using growth factors. The collagencomponent of the artificial skin often contracts to less than one tenthof its original area when populated by fibroblasts.

Cicatricial contraction, contraction due to shrinkage of the fibroustissue of a scar, is common. In some cases the scar may become a viciouscicatrix, a scar in which the contraction causes serious deformity. Apatient's stomach may be effectively separated into two separatechambers in an hour-glass contracture by the contraction of scar tissueformed when a stomach ulcer heals. Obstruction of passages and ducts,cicatricial stenosis, may occur due to the contraction of scar tissue.Contraction of blood vessels may be due to primary obstruction orsurgical trauma, for example, after surgery or angioplasty. Stenosis ofother hollow visci, for examples, ureters, may also occur. Problems mayoccur where any form of scarring takes place, whether resulting fromaccidental wounds or from surgery. Conditions of the skin and tendonswhich involve contraction of collagen-including tissues includepost-trauma conditions resulting from surgery or accidents, for example,hand or foot tendon injuries, post-graft conditions and pathologicalconditions, such as scleroderma, Dupuytren's contracture andepidermolysis bullosa. Scarring and contraction of tissues in the eyemay occur in various conditions, for example, the sequelae of retinaldetachment or diabetic eye disease (as mentioned above). Contraction ofthe sockets found in the skull for the eyeballs and associatedstructures, including extra-ocular muscles and eyelids, may occur ifthere is trauma or inflammatory damage. The tissues contract within thesockets causing a variety of problems including double vision and anunsightly appearance.

For further information on different types of fibrosis see: Molina V, etal., (2002), “Fibrotic diseases”, Harefuah, 141(11): 973-8, 1009; Yu L,et al., (2002), “Therapeutic strategies to halt renal fibrosis”, CurrOpin Pharmacol. 2(2):177-81; Keane WF and Lyle P A. (2003), “Recentadvances in management of type 2 diabetes and nephropathy: lessons fromthe RENAAL study”, Am J Kidney Dis. 41(3 Suppl 2): S22-5; Bohle A, etal., (1989), “The pathogenesis of chronic renal failure”, Pathol ResPract. 185(4):421-40; Kikkawa R, et al., (1997), “Mechanism of theprogression of diabetic nephropathy to renal failure”, Kidney Int Suppl.62:S39-40; Bataller R, and Brenner D A. (2001), “Hepatic stellate cellsas a target for the treatment of liver fibrosis”, Semin Liver Dis.21(3):437-51; Gross T J and Hunninghake G W, (2001) “Idiopathicpulmonary fibrosis”, N Engl J. Med. 345(7):517-25; Frohlich E D. (2001)“Fibrosis and ischemia: the real risks in hypertensive heart disease”,Am J Hypertens; 14(6 Pt 2):194S-199S; Friedman SL. (2003), “Liverfibrosis—from bench to bedside”, J. Hepatol. 38 Suppl 1:S38-53; AlbanisE, et al., (2003), “Treatment of hepatic fibrosis: almost there”, CurrGastroenterol Rep. 5(1):48-56; (Weber K T. (2000), “Fibrosis andhypertensive heart disease”, Curr Opin Cardiol. 15(4):264-72).

Delivery of Nucleic Acid Molecules and Pharmaceutical Formulations

Nucleic acid molecules may be adapted for use to prevent or treatfibroses (e.g., liver, kidney, peritoneal, and pulmonary) diseases,traits, conditions and/or disorders, and/or any other trait, disease,disorder or condition that is related to or will respond to the levelsof hsp47 in a cell or tissue, alone or in combination with othertherapies. A nucleic acid molecule may include a delivery vehicle,including liposomes, for administration to a subject, carriers anddiluents and their salts, and/or can be present in pharmaceuticallyacceptable formulations.

Nucleic acid molecules disclosed herein may be delivered or administereddirectly with a carrier or diluent but not any delivery vehicle thatacts to assist, promote or facilitate entry to the cell, including viralvectors, viral particles, liposome formulations, lipofectin orprecipitating agents and the like.

Nucleic acid molecules may be delivered or administered to a subject bydirect application of the nucleic acid molecules with a carrier ordiluent or any other delivery vehicle that acts to assist, promote orfacilitate entry into a cell, including viral sequences, viralparticular, liposome formulations, lipofectin or precipitating agentsand the like. Polypeptides that facilitate introduction of nucleic acidinto a desired subject such as those described in US. ApplicationPublication No. 20070155658 (e.g., a melamine derivative such as2,4,6-Triguanidino Traizine and 2,4,6-Tramidosarcocyl Melamine, apolyarginine polypeptide, and a polypeptide including alternatingglutamine and asparagine residues).

Methods for the delivery of nucleic acid molecules are described inAkhtar et al., Trends Cell Bio., 2: 139 (1992); Delivery Strategies forAntisense Oligonucleotide Therapeutics, ed. Akhtar, (1995), Maurer etal., Mol. Membr. Biol., 16: 129-140 (1999); Hofland and Huang, Handb.Exp. Pharmacol., 137: 165-192 (1999); and Lee et al., ACS Symp. Ser.,752: 184-192 (2000); U.S. Pat. Nos. 6,395,713; 6,235,310; 5,225,182;5,169,383; 5,167,616; 4,959217; 4,925,678; 4,487,603; and 4,486,194 andSullivan et al., PCT WO 94/02595; PCT WO 00/03683 and PCT WO 02/08754;and U.S. Patent Application Publication No. 2003077829. These protocolscan be utilized for the delivery of virtually any nucleic acid molecule.Nucleic acid molecules can be administered to cells by a variety ofmethods known to those of skill in the art, including, but notrestricted to, encapsulation in liposomes, by iontophoresis, or byincorporation into other vehicles, such as biodegradable polymers,hydrogels, cyclodextrins (see e.g., Gonzalez et al., Bioconjugate Chem.,10: 1068-1074 (1999); Wang et al., International PCT publication Nos. WO03/47518 and WO 03/46185), poly(lactic-co-glycolic)acid (PLGA) and PLCAmicrospheres (see for example U.S. Pat. No. 6,447,796 and U.S.Application Publication No. 2002130430), biodegradable nanocapsules, andbioadhesive microspheres, or by proteinaceous vectors (O'Hare andNormand, International PCT Publication No. WO 00/53722). Alternatively,the nucleic acid/vehicle combination is locally delivered by directinjection or by use of an infusion pump. Direct injection of the nucleicacid molecules of the invention, whether subcutaneous, intramuscular, orintradermal, can take place using standard needle and syringemethodologies, or by needle-free technologies such as those described inConry et al., Clin. Cancer Res., 5: 2330-2337 (1999) and Barry et al.,International PCT Publication No. WO 99/31262. The molecules of theinstant invention can be used as pharmaceutical agents. Pharmaceuticalagents prevent, modulate the occurrence, or treat (alleviate a symptomto some extent, preferably all of the symptoms) of a disease state in asubject.

Nucleic acid molecules may be complexed with cationic lipids, packagedwithin liposomes, or otherwise delivered to target cells or tissues. Thenucleic acid or nucleic acid complexes can be locally administered torelevant tissues ex vivo, or in vivo through direct dermal application,transdermal application, or injection, with or without theirincorporation in biopolymers. The nucleic acid molecules of theinvention may include sequences shown in Tables I. Examples of suchnucleic acid molecules consist essentially of sequences provided inTable I.

Delivery systems include surface-modified liposomes containingpoly(ethylene glycol) lipids (PEG-modified, or long-circulatingliposomes or stealth liposomes). These formulations offer a method forincreasing the accumulation of drugs in target tissues. This class ofdrug carriers resists opsonization and elimination by the mononuclearphagocytic system (MPS or RES), thereby enabling longer bloodcirculation times and enhanced tissue exposure for the encapsulated drug(Lasic et al. Chem. Rev. 1995, 95, 2601-2627; Ishiwata et al., Chem.Pharm. Bull. 1995, 43, 1005-1011).

Nucleic acid molecules may be formulated or complexed withpolyethylenimine (e.g., linear or branched PEI) and/or polyethyleniminederivatives, including for examplepolyethyleneimine-polyethyleneglycol-N-acetylgalactosamine (PEI-PEG-GAL)or polyethyleneimine-polyethyleneglycol-tri-N-acetylgalactosamine(PEI-PEG-triGAL) derivatives, grafted PEIs such as galactose PEI,cholesterol PEI, antibody derivatized PEI, and polyethylene glycol PEI(PEG-PEI) derivatives thereof (see for example Ogris et al., 2001, AAPAPharmSci, 3, 1-11; Furgeson et al., 2003, Bioconjugate Chem., 14,840-847; Kunath et al., 2002, Pharmaceutical Research, 19, 810-817; Choiet al, 2001, Bull. Korean Chem. Soc., 22, 46-52; Bettinger et al., 1999,Bioconjugate Chem., 10, 558-561; Peterson et al., 2002, BioconjugateChem., 13, 845-854; Erbacher et al., 1999, Journal of Gene MedicinePreprint, 1, 1-18; Godbey et al., 1999, PNAS USA, 96, 5177-5181; Godbeyet al., 1999, Journal of Controlled Release, 60, 149-160; Diebold etal., 1999, Journal of Biological Chemistry, 274, 19087-19094; Thomas andKlibanov, 2002, PNAS USA, 99, 14640-14645; Sagara, U.S. Pat. No.6,586,524 and United States Patent Application Publication No.20030077829.

Nucleic acid molecules may be complexed with membrane disruptive agentssuch as those described in U.S. Patent Application Publication No.20010007666. The membrane disruptive agent or agents and the nucleicacid molecule may also be complexed with a cationic lipid or helperlipid molecule, such as those lipids described in U.S. Pat. No.6,235,310.

The nucleic acid molecules may be administered via pulmonary delivery,such as by inhalation of an aerosol or spray dried formulationadministered by an inhalation device or nebulizer, providing rapid localuptake of the nucleic acid molecules into relevant pulmonary tissues.Solid particulate compositions containing respirable dry particles ofmicronized nucleic acid compositions can be prepared by grinding driedor lyophilized nucleic acid compositions, and then passing themicronized composition through, for example, a 400 mesh screen to breakup or separate out large agglomerates. A solid particulate compositioncomprising the nucleic acid compositions of the invention can optionallycontain a dispersant which serves to facilitate the formation of anaerosol as well as other therapeutic compounds. A suitable dispersant islactose, which can be blended with the nucleic acid compound in anysuitable ratio, such as a 1 to 1 ratio by weight.

Aerosols of liquid particles may include a nucleic acid moleculesdisclosed herein and can be produced by any suitable means, such as witha nebulizer (see e.g., U.S. Pat. No. 4,501,729). Nebulizers arecommercially available devices which transform solutions or suspensionsof an active ingredient into a therapeutic aerosol mist either by meansof acceleration of a compressed gas, typically air or oxygen, through anarrow venturi orifice or by means of ultrasonic agitation. Suitableformulations for use in nebulizers include the active ingredient in aliquid carrier in an amount of up to 40% w/w preferably less than 20%w/w of the formulation. The carrier is typically water or a diluteaqueous alcoholic solution, preferably made isotonic with body fluids bythe addition of, e.g., sodium chloride or other suitable salts. Optionaladditives include preservatives if the formulation is not preparedsterile, e.g., methyl hydroxybenzoate, anti-oxidants, flavorings,volatile oils, buffering agents and emulsifiers and other formulationsurfactants. The aerosols of solid particles including the activecomposition and surfactant can likewise be produced with any solidparticulate aerosol generator. Aerosol generators for administeringsolid particulate therapeutics to a subject produce particles which arerespirable, as explained above, and generate a volume of aerosolcontaining a predetermined metered dose of a therapeutic composition ata rate suitable for human administration. One illustrative type of solidparticulate aerosol generator is an insufflator. Suitable formulationsfor administration by insufflation include finely comminuted powderswhich can be delivered by means of an insufflator. In the insufflator,the powder, e.g., a metered dose thereof effective to carry out thetreatments described herein, is contained in capsules or cartridges,typically made of gelatin or plastic, which are either pierced or openedin situ and the powder delivered by air drawn through the device uponinhalation or by means of a manually-operated pump. The powder employedin the insufflator consists either solely of the active ingredient or ofa powder blend comprising the active ingredient, a suitable powderdiluent, such as lactose, and an optional surfactant. The activeingredient typically includes from 0.1 to 100 w/w of the formulation. Asecond type of illustrative aerosol generator includes a metered doseinhaler. Metered dose inhalers are pressurized aerosol dispensers,typically containing a suspension or solution formulation of the activeingredient in a liquefied propellant. During use these devices dischargethe formulation through a valve adapted to deliver a metered volume toproduce a fine particle spray containing the active ingredient. Suitablepropellants include certain chlorofluorocarbon compounds, e.g.,dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane and mixtures thereof. The formulation canadditionally contain one or more co-solvents, for example, ethanol,emulsifiers and other formulation surfactants, such as oleic acid orsorbitan trioleate, anti-oxidants and suitable flavoring agents. Othermethods for pulmonary delivery are described in, e.g., US PatentApplication No. 20040037780, and U.S. Pat. Nos. 6,592,904; 6,582,728;6,565,885. PCT Patent Publication No. WO2008/132723 relates to aerosoldelivery of oligonucleotides in general, and of siRNA in particular, tothe respiratory system.

Nucleic acid molecules may be administered to the central nervous system(CNS) or peripheral nervous system (PNS). Experiments have demonstratedthe efficient in vivo uptake of nucleic acids by neurons. See e.g.,Sommer et al., 1998, Antisense Nuc. Acid Drug Dev., 8, 75; Epa et al.,2000, Antisense Nuc. Acid Drug Dev., 10, 469; Broaddus et al., 1998, J.Neurosurg., 88(4), 734; Karle et al., 1997, Eur. J. Pharmocol.,340(2/3), 153; Bannai et al., 1998, Brain Research, 784(1,2), 304;Rajakumar et al., 1997, Synapse, 26(3), 199; Wu-pong et al., 1999,BioPharm, 12(1), 32; Bannai et al., 1998, Brain Res. Protoc., 3(1), 83;and Simantov et al., 1996, Neuroscience, 74(1), 39. Nucleic acidmolecules are therefore amenable to delivery to and uptake by cells inthe CNS and/or PNS.

Delivery of nucleic acid molecules to the CNS is provided by a varietyof different strategies. Traditional approaches to CNS delivery that canbe used include, but are not limited to, intrathecal andintracerebroventricular administration, implantation of catheters andpumps, direct injection or perfusion at the site of injury or lesion,injection into the brain arterial system, or by chemical or osmoticopening of the blood-brain barrier. Other approaches can include the useof various transport and carrier systems, for example though the use ofconjugates and biodegradable polymers. Furthermore, gene therapyapproaches, e.g., as described in Kaplitt et al., U.S. Pat. No.6,180,613 and Davidson, WO 04/013280, can be used to express nucleicacid molecules in the CNS.

Delivery systems may include, for example, aqueous and nonaqueous gels,creams, multiple emulsions, microemulsions, liposomes, ointments,aqueous and nonaqueous solutions, lotions, aerosols, hydrocarbon basesand powders, and can contain excipients such as solubilizers, permeationenhancers (e.g., fatty acids, fatty acid esters, fatty alcohols andamino acids), and hydrophilic polymers (e.g., polycarbophil andpolyvinylpyrolidone). In one embodiment, the pharmaceutically acceptablecarrier is a liposome or a transdermal enhancer. Examples of liposomeswhich can be used in this invention include the following: (1)CellFectin, 1:1.5 (M/M) liposome formulation of the cationic lipidN,NI,NII,NIII-tetramethyl-N,NI,NII,NIII-tetrapalmit-y-spermine anddioleoyl phosphatidylethanolamine (DOPE) (GIBCO BRL); (2) CytofectinGSV, 2:1 (M/M) liposome formulation of a cationic lipid and DOPE (GlenResearch); (3)DOTAP(N-[1-(2,3-dioleoyloxy)-N,N,N-tri-methyl-ammoniummethylsulfate)(Boehringer Manheim); and (4) Lipofectamine, 3:1 (M/M) liposomeformulation of the polycationic lipid DOSPA, the neutral lipid DOPE(GIBCO BRL) and Di-Alkylated Amino Acid (DiLA2).

Delivery systems may include patches, tablets, suppositories, pessaries,gels and creams, and can contain excipients such as solubilizers andenhancers (e.g., propylene glycol, bile salts and amino acids), andother vehicles (e.g., polyethylene glycol, fatty acid esters andderivatives, and hydrophilic polymers such ashydroxypropylmethylcellulose and hyaluronic acid).

Nucleic acid molecules may be formulated or complexed withpolyethylenimine (e.g., linear or branched PEI) and/or polyethyleniminederivatives, including for example grafted PEIs such as galactose PEI,cholesterol PEI, antibody derivatized PEI, and polyethylene glycol PEI(PEG-PEI) derivatives thereof (see for example Ogris et al., 2001, AAPAPharmSci, 3, 1-11; Furgeson et al., 2003, Bioconjugate Chem., 14,840-847; Kunath et al., 2002, Pharmaceutical Research, 19, 810-817; Choiet al., 2001, Bull. Korean Chem. Soc., 22, 46-52; Bettinger et al.,1999, Bioconjugate Chem., 10, 558-561; Peterson et al., 2002,Bioconjugate Chem., 13, 845-854; Erbacher et al., 1999, Journal of GeneMedicine Preprint, 1, 1-18; Godbey et al., 1999., PNAS USA, 96,5177-5181; Godbey et al., 1999, Journal of Controlled Release, 60,149-160; Diebold et al., 1999, Journal of Biological Chemistry, 274,19087-19094; Thomas and Klibanov, 2002, PNAS USA, 99, 14640-14645; andSagara, U.S. Pat. No. 6,586,524.

Nucleic acid molecules may include a bioconjugate, for example a nucleicacid conjugate as described in Vargeese et al., U.S. Ser. No.10/427,160; U.S. Pat. No. 6,528,631; U.S. Pat. No. 6,335,434; U.S. Pat.No. 6,235,886; U.S. Pat. No. 6,153,737; U.S. Pat. No. 5,214,136; U.S.Pat. No. 5,138,045.

Compositions, methods and kits disclosed herein may include anexpression vector that includes a nucleic acid sequence encoding atleast one nucleic acid molecule of the invention in a manner that allowsexpression of the nucleic acid molecule. Methods of introducing nucleicacid molecules or one or more vectors capable of expressing the strandsof dsRNA into the environment of the cell will depend on the type ofcell and the make up of its environment. The nucleic acid molecule orthe vector construct may be directly introduced into the cell (i.e.,intracellularly); or introduced extracellularly into a cavity,interstitial space, into the circulation of an organism, introducedorally, or may be introduced by bathing an organism or a cell in asolution containing dsRNA. The cell is preferably a mammalian cell; morepreferably a human cell. The nucleic acid molecule of the expressionvector can include a sense region and an antisense region. The antisenseregion can include a sequence complementary to a RNA or DNA sequenceencoding hsp47 and the sense region can include a sequence complementaryto the antisense region. The nucleic acid molecule can include twodistinct strands having complementary sense and antisense regions. Thenucleic acid molecule can include a single strand having complementarysense and antisense regions.

Nucleic acid molecules that interact with target RNA molecules anddown-regulate gene encoding target RNA molecules (e.g., target RNAmolecules referred to by Genbank Accession numbers herein) may beexpressed from transcription units inserted into DNA or RNA vectors.Recombinant vectors can be DNA plasmids or viral vectors. Nucleic acidmolecule expressing viral vectors can be constructed based on, but notlimited to, adeno-associated virus, retrovirus, adenovirus, oralphavirus. The recombinant vectors capable of expressing the nucleicacid molecules can be delivered as described herein, and persist intarget cells. Alternatively, viral vectors can be used that provide fortransient expression of nucleic acid molecules. Such vectors can berepeatedly administered as necessary. Once expressed, the nucleic acidmolecules bind and down-regulate gene function or expression via RNAinterference (RNAi). Delivery of nucleic acid molecule expressingvectors can be systemic, such as by intravenous or intramuscularadministration, by administration to target cells ex-planted from asubject followed by reintroduction into the subject, or by any othermeans that would allow for introduction into the desired target cell.

Expression vectors may include a nucleic acid sequence encoding at leastone nucleic acid molecule disclosed herein, in a manner which allowsexpression of the nucleic acid molecule. For example, the vector maycontain sequence(s) encoding both strands of a nucleic acid moleculethat include a duplex. The vector can also contain sequence(s) encodinga single nucleic acid molecule that is self-complementary and thus formsa nucleic acid molecule. Non-limiting examples of such expressionvectors are described in Paul et al., 2002, Nature Biotechnology, 19,505; Miyagishi and Taira, 2002, Nature Biotechnology, 19, 497; Lee etal., 2002, Nature Biotechnology, 19, 500; and Novina et al., 2002,Nature Medicine, advance online publication doi:10.1038/nm725.Expression vectors may also be included in a mammalian (e.g., human)cell.

An expression vector may include a nucleic acid sequence encoding two ormore nucleic acid molecules, which can be the same or different.Expression vectors may include a sequence for a nucleic acid moleculecomplementary to a nucleic acid molecule referred to by a GenbankAccession number NM_(—)001235, for example those shown in Table I.

An expression vector may encode one or both strands of a nucleic acidduplex, or a single self-complementary strand that self hybridizes intoa nucleic acid duplex. The nucleic acid sequences encoding nucleic acidmolecules can be operably linked in a manner that allows expression ofthe nucleic acid molecule (see for example Paul et al., 2002, NatureBiotechnology, 19, 505; Miyagishi and Taira, 2002, Nature Biotechnology,19, 497; Lee et al., 2002, Nature Biotechnology, 19, 500; and Novina etal., 2002, Nature Medicine, advance online publicationdoi:10.1038/nm725).

An expression vector may include one or more of the following: a) atranscription initiation region (e.g., eukaryotic pol I, II or IIIinitiation region); b) a transcription termination region (e.g.,eukaryotic pol I, II or III termination region); c) an intron and d) anucleic acid sequence encoding at least one of the nucleic acidmolecules, wherein said sequence is operably linked to the initiationregion and the termination region in a manner that allows expressionand/or delivery of the nucleic acid molecule. The vector can optionallyinclude an open reading frame (ORF) for a protein operably linked on the5′ side or the 3′-side of the sequence encoding the nucleic acidmolecule; and/or an intron (intervening sequences).

Transcription of the nucleic acid molecule sequences can be driven froma promoter for eukaryotic RNA polymerase I (pol I), RNA polymerase II(pol II), or RNA polymerase III (pol III). Transcripts from pol II orpol III promoters are expressed at high levels in all cells; the levelsof a given pol II promoter in a given cell type depends on the nature ofthe gene regulatory sequences (enhancers, silencers, etc.) presentnearby. Prokaryotic RNA polymerase promoters are also used, providingthat the prokaryotic RNA polymerase enzyme is expressed in theappropriate cells (Elroy-Stein and Moss, 1990, Proc. Natl. Acad. Sci.USA, 87, 6743-7; Gao and Huang 1993, Nucleic Acids Res., 21, 2867-72;Lieber et al., 1993, Methods Enzymol., 217, 47-66; Zhou et al., 1990,Mol. Cell. Biol., 10, 4529-37). Several investigators have demonstratedthat nucleic acid molecules expressed from such promoters can functionin mammalian cells (e.g. Kashani-Sabet et al., 1992, Antisense Res.Dev., 2, 3-15; Ojwang et al., 1992, Proc. Natl. Acad. Sci. USA, 89,10802-6; Chen et al., 1992, Nucleic Acids Res., 20, 4581-9; Yu et al.,1993, Proc. Natl. Acad. Sci. USA, 90, 6340-4; L′Huillier et al., 1992,EMBO J., 11, 4411-8; Lisziewicz et al., 1993, Proc. Natl. Acad. Sci.U.S.A, 90, 8000-4; Thompson et al., 1995, Nucleic Acids Res., 23, 2259;Sullenger & Cech, 1993, Science, 262, 1566). More specifically,transcription units such as the ones derived from genes encoding U6small nuclear (snRNA), transfer RNA (tRNA) and adenovirus VA RNA areuseful in generating high concentrations of desired RNA molecules suchas siNA in cells (Thompson et al., supra; Couture and Stinchcomb, 1996,supra; Noonberg et al., 1994, Nucleic Acid Res., 22, 2830; Noonberg etal., U.S. Pat. No. 5,624,803; Good et al., 1997, Gene Ther., 4, 45;Beigelman et al., International PCT Publication No. WO 96/18736. Theabove nucleic acid transcription units can be incorporated into avariety of vectors for introduction into mammalian cells, including butnot restricted to, plasmid DNA vectors, viral DNA vectors (such asadenovirus or adeno-associated virus vectors), or viral RNA vectors(such as retroviral or alphavirus vectors) (see Couture and Stinchcomb,1996 supra).

Nucleic acid molecule may be expressed within cells from eukaryoticpromoters (e.g., Izant and Weintraub, 1985, Science, 229, 345; McGarryand Lindquist, 1986, Proc. Natl. Acad. Sci., USA 83, 399; Scanlon etal., 1991, Proc. Natl. Acad. Sci. USA, 88, 10591-5; Kashani-Sabet etal., 1992, Antisense Res. Dev., 2, 3-15; propulic et al., 1992, J.Virol., 66, 1432-41; Weerasinghe et al., 1991, J. Virol., 65, 5531-4;Ojwang et al., 1992, Proc. Natl. Acad. Sci. USA, 89, 10802-6; Chen etal., 1992, Nucleic Acids Res. 20, 4581-9; Sarver et al., 1990 Science,247, 1222-1225; Thompson et al., 1995, Nucleic Acids Res., 23, 2259;Good et al., 1997, Gene Therapy, 4, 45. Those skilled in the art realizethat any nucleic acid can be expressed in eukaryotic cells from theappropriate DNA/RNA vector. The activity of such nucleic acids can beaugmented by their release from the primary transcript by a enzymaticnucleic acid (Draper et al., PCT WO 93/23569, and Sullivan et al., PCTWO 94/02595; Ohkawa et al., 1992, Nucleic Acids Symp. Ser., 27, 15-6;Taira et al., 1991, Nucleic Acids Res., 19, 5125-30; Ventura et al.,1993, Nucleic Acids Res., 21, 3249-55; Chowrira et al., 1994, J. Biol.Chem., 269, 25856.

A viral construct packaged into a viral particle would accomplish bothefficient introduction of an expression construct into the cell andtranscription of dsRNA construct encoded by the expression construct.

Methods for oral introduction include direct mixing of RNA with food ofthe organism, as well as engineered approaches in which a species thatis used as food is engineered to express an RNA, then fed to theorganism to be affected. Physical methods may be employed to introduce anucleic acid molecule solution into the cell. Physical methods ofintroducing nucleic acids include injection of a solution containing thenucleic acid molecule, bombardment by particles covered by the nucleicacid molecule, soaking the cell or organism in a solution of the RNA, orelectroporation of cell membranes in the presence of the nucleic acidmolecule.

Other methods known in the art for introducing nucleic acids to cellsmay be used, such as lipid-mediated carrier transport, chemical mediatedtransport, such as calcium phosphate, and the like. Thus the nucleicacid molecules may be introduced along with components that perform oneor more of the following activities: enhance RNA uptake by the cell,promote annealing of the duplex strands, stabilize the annealed strands,or other-wise increase inhibition of the target gene.

The nucleic acid molecules or the vector construct can be introducedinto the cell using suitable formulations. One preferable formulation iswith a lipid formulation such as in Lipofectamine™ 2000 (Invitrogen, CA,USA), vitamin A coupled liposomes (Sato et al. Nat Biotechnol 2008;26:431-442, PCT Patent Publication No. WO 2006/068232). Lipidformulations can also be administered to animals such as by intravenous,intramuscular, or intraperitoneal injection, or orally or by inhalationor other methods as are known in the art. When the formulation issuitable for administration into animals such as mammals and morespecifically humans, the formulation is also pharmaceuticallyacceptable. Pharmaceutically acceptable formulations for administeringoligonucleotides are known and can be used. In some instances, it may bepreferable to formulate dsRNA in a buffer or saline solution anddirectly inject the formulated dsRNA into cells, as in studies withoocytes. The direct injection of dsRNA duplexes may also be done. Forsuitable methods of introducing dsRNA see U.S. published patentapplication No. 2004/0203145, 20070265220 which are incorporated hereinby reference.

Polymeric nanocapsules or microcapsules facilitate transport and releaseof the encapsulated or bound dsRNA into the cell. They include polymericand monomeric materials, especially including polybutylcyanoacrylate. Asummary of materials and fabrication methods has been published (seeKreuter, 1991). The polymeric materials which are formed from monomericand/or oligomeric precursors in the polymerization/nanoparticlegeneration step, are per se known from the prior art, as are themolecular weights and molecular weight distribution of the polymericmaterial which a person skilled in the field of manufacturingnanoparticles may suitably select in accordance with the usual skill.

Nucleic acid moles may be formulated as a microemulsion. A microemulsionis a system of water, oil and amphiphile which is a single opticallyisotropic and thermodynamically stable liquid solution. Typicallymicroemulsions are prepared by first dispersing an oil in an aqueoussurfactant solution and then adding a sufficient amount of a 4thcomponent, generally an intermediate chain-length alcohol to form atransparent system.

Surfactants that may be used in the preparation of microemulsionsinclude, but are not limited to, ionic surfactants, non-ionicsurfactants, Brij 96, polyoxyethylene oleyl ethers, polyglycerol fattyacid esters, tetraglycerol monolaurate (ML310), tetraglycerol monooleate(MO310), hexaglycerol monooleate (PO310), hexaglycerol pentaoleate(PO500), decaglycerol monocaprate (MCA750), decaglycerol monooleate(MO750), decaglycerol sequioleate (SO750), decaglycerol decaoleate(DA0750), alone or in combination with cosurfactants. The cosurfactant,usually a short-chain alcohol such as ethanol, 1-propanol, and1-butanol, serves to increase the interfacial fluidity by penetratinginto the surfactant film and consequently creating a disordered filmbecause of the void space generated among surfactant molecules.

Water Soluble Crosslinked Polymers

Delivery formulations can include water soluble degradable crosslinkedpolymers that include one or more degradable crosslinking lipid moiety,one or more PEI moiety, and/or one or more mPEG (methyl ether derivativeof PEG (tnethoxypoly(ethylene glycol)).

Degradable lipid moieties preferably include compounds having thefollowing structural motif:

In the above formula, ester linkages are biodegradable groups, Rrepresents a relatively hydrophobic “lipo” group, and the structuralmotif shown occurs m times where m is in the range of about 1 to about30. For example, in certain embodiments R is selected from the groupconsisting of C2-C50 alkyl, C2-C50 heteroalkyl, C2-C50 alkenyl, C2-C50heteroalkenyl, C5-C50 aryl; C2-C50 heteroaryl; C2-C50 alkynyl, C2-C50heteroalkynyl, C2-C50 carboxyalkenyl, and C2-C50 carboxyheteroalkenyl.In preferred embodiments. R is a saturated or unsaturated alkyl having 4to 30 carbons, more preferably 8 to 24 carbons or a sterol, preferably acholesteryl moiety. In preferred embodiments, R is oleic, lauric,myristic, palmitic margaric, stearic, arachidic, behenic, or lignoceric.In a most preferred embodiment, R is oleic.

The N in formula (B) may have an electron pair or a bond to a hydrogenatom. When N has an electron pair, the recurring unit may be cationic atlow pH.

The degradable crosslinking lipid moiety may be reacted with apolyethyleneimine (PEI) as shown in Scheme A below:

In formula (A), R has the same meanings as described above. The PEI maycontain recurring units of formula (B) in which x is an integer in therange of about 1 to about 100 and y is an integer in the range of about1 to about 100.

The reaction illustrated in Scheme A may be carried out by intermixingthe PEI and the diacrylate (I) in a mutual solvent such as ethanol,methanol or dichloromethane with stirring, preferably at roomtemperature for several hours, then evaporating the solvent to recoverthe resulting polymer. While not wishing to be bound to any particulartheory, it is believed that the reaction between the PEI and diacrylate(I) involves a Michael reaction between one or more amines of the PEIwith double bond(s) of the diacrylate (see J. March, Advanced OrganicChemistry 3_(rd) Ed, pp. 711-712 (1985)). The di acrylate shown inScheme A may be prepared in the manner as described in U.S. applicationSer. No. 11/216,986 (US Publication No. 2006/0258751).

The molecular weight of the PEI is preferably in the range of about 200to 25,000 Daltons more preferably 400 to 5,000 Daltons, yet morepreferably 600 to 2000 Daltons. PEI may be either branched or linear.

The molar ratio of PEI to diacrylate is preferably in the range of about1:2 to about 1:20. The weight average molecular weight of the cationiclipopolymer may be in the range of about 500 Daltons to about 1,000,000Daltons preferably in the range of about 2,000 Daltons to about 200.000Daltons. Molecular weights may be determined by size exclusionchromatography using PEG standards or by agarose gel electrophoresis.

The cationic lipopolymer is preferably degradable, more preferablybiodegradable, e.g., degradable by a mechanism selected from the groupconsisting of hydrolysis, enzyme cleavage, reduction, photo-cleavage,and sonication. While not wishing to be bound to any particular theory,but it is believed that degradation of the cationic lipopolymer offormula (II) within the cell proceeds by enzymatic cleavage and/orhydrolysis of the ester linkages.

Synthesis may be carried out by reacting the degradable lipid moietywith the PEI moiety as described above. Then the mPEG (methyl etherderivative of PEG (methoxypoly (ethylene glycol)), is added to form thedegradable crosslinked polymer. In preferred embodiments, the reactionis carried out at room temperature. The reaction products may beisolated by any means known in the art including chromatographictechniques. In a preferred embodiment, the reaction product may beremoved by precipitation followed by centrifugation.

Dosages

The useful dosage to be administered and the particular mode ofadministration will vary depending upon such factors as the cell type,or for in vivo use, the age, weight and the particular animal and regionthereof to be treated, the particular nucleic acid and delivery methodused, the therapeutic or diagnostic use contemplated, and the form ofthe formulation, for example, suspension, emulsion, micelle or liposome,as will be readily apparent to those skilled in the art. Typically,dosage is administered at lower levels and increased until the desiredeffect is achieved.

When lipids are used to deliver the nucleic acid, the amount of lipidcompound that is administered can vary and generally depends upon theamount of nucleic acid being administered. For example, the weight ratioof lipid compound to nucleic acid is preferably from about 1:1 to about30:1, with a weight ratio of about 5:1 to about 10:1 being morepreferred.

A suitable dosage unit of nucleic acid molecules may be in the range of0.001 to 0.25 milligrams per kilogram body weight of the recipient perday, or in the range of 0.01 to 20 micrograms per kilogram body weightper day, or in the range of 0.01 to 10 micrograms per kilogram bodyweight per day, or in the range of 0.10 to 5 micrograms per kilogrambody weight per day, or in the range of 0.1 to 2.5 micrograms perkilogram body weight per day.

Suitable amounts of nucleic acid molecules may be introduced and theseamounts can be empirically determined using standard methods. Effectiveconcentrations of individual nucleic acid molecule species in theenvironment of a cell may be about 1 femtomolar, about 50 femtomolar,100 femtomolar, 1 picomolar, 1.5 picomolar, 2.5 picomolar, 5 picomolar,10 picomolar, 25 picomolar, 50 picomolar, 100 picomolar, 500 picomolar,1 nanomolar, 2.5 nanomolar, 5 nanomolar, 10 nanomolar, 25 nanomolar, 50nanomolar, 100 nanomolar, 500 nanomolar, 1 micromolar, 2.5 micromolar, 5micromolar, 10 micromolar, 100 micromolar or more.

Dosage may be from 0.01 □g to 1 g per kg of body weight (e.g., 0.1 □g,0.25 □g, 0.5 □g, 0.75 □g, 1 □g, 2.5 □g, 5 □g, 10 □g, 25 □g, 50 □g, 100□g, 250 □g, 500 □g, 1 mg, 2.5 mg, 5 mg, 10 mg, 25 mg, 50 mg, 100 mg, 250mg, or 500 mg per kg).

Dosage levels of the order of from about 0.1 mg to about 140 mg perkilogram of body weight per day are useful in the treatment of theabove-indicated conditions (about 0.5 mg to about 7 g per subject perday). The amount of active ingredient that can be combined with thecarrier materials to produce a single dosage form varies depending uponthe host treated and the particular mode of administration. Dosage unitforms generally contain between from about 1 mg to about 500 mg of anactive ingredient.

It is understood that the specific dose level for any particular subjectdepends upon a variety of factors including the activity of the specificcompound employed, the age, body weight, general health, sex, diet, timeof administration, route of administration, and rate of excretion, drugcombination and the severity of the particular disease undergoingtherapy.

Pharmaceutical compositions that include the nucleic acid moleculedisclosed herein may be administered once daily, qid, tid, bid, QD, orat any interval and for any duration that is medically appropriate.However, the therapeutic agent may also be dosed in dosage unitscontaining two, three, four, five, six or more sub-doses administered atappropriate intervals throughout the day. In that case, the nucleic acidmolecules contained in each sub-dose may be correspondingly smaller inorder to achieve the total daily dosage unit. The dosage unit can alsobe compounded for a single dose over several days, e.g., using aconventional sustained release formulation which provides sustained andconsistent release of the dsRNA over a several day period. Sustainedrelease formulations are well known in the art. The dosage unit maycontain a corresponding multiple of the daily dose. The composition canbe compounded in such a way that the sum of the multiple units of anucleic acid together contain a sufficient dose.

Pharmaceutical Compositions, Kits, and Containers

Also provided are compositions, kits, containers and formulations thatinclude a nucleic acid molecule (e.g., an siNA molecule) as providedherein for reducing expression of hsp47 for administering ordistributing the nucleic acid molecule to a patient. A kit may includeat least one container and at least one label. Suitable containersinclude, for example, bottles, vials, syringes, and test tubes. Thecontainers can be formed from a variety of materials such as glass,metal or plastic. The container can hold amino acid sequence(s), smallmolecule(s), nucleic acid sequence(s), cell population(s) and/orantibody(s). In one embodiment, the container holds a polynucleotide foruse in examining the mRNA expression profile of a cell, together withreagents used for this purpose. In another embodiment a containerincludes an antibody, binding fragment thereof or specific bindingprotein for use in evaluating hsp47 protein expression cells andtissues, or for relevant laboratory, prognostic, diagnostic,prophylactic and therapeutic purposes; indications and/or directions forsuch uses can be included on or with such container, as can reagents andother compositions or tools used for these purposes. Kits may furtherinclude associated indications and/or directions; reagents and othercompositions or tools used for such purpose can also be included.

The container can alternatively hold a composition that is effective fortreating, diagnosis, prognosing or prophylaxing a condition and can havea sterile access port (for example the container can be an intravenoussolution bag or a vial having a stopper pierceable by a hypodermicinjection needle). The active agents in the composition can be a nucleicacid molecule capable of specifically binding hsp47 and/or modulatingthe function of hsp47.

A kit may further include a second container that includes apharmaceutically-acceptable buffer, such as phosphate-buffered saline,Ringer's solution and/or dextrose solution. It can further include othermaterials desirable from a commercial and user standpoint, includingother buffers, diluents, filters, stirrers, needles, syringes, and/orpackage inserts with indications and/or instructions for use.

The units dosage ampoules or multidose containers, in which the nucleicacid molecules are packaged prior to use, may include an hermeticallysealed container enclosing an amount of polynucleotide or solutioncontaining a polynucleotide suitable for a pharmaceutically effectivedose thereof, or multiples of an effective dose. The polynucleotide ispackaged as a sterile formulation, and the hermetically sealed containeris designed to preserve sterility of the formulation until use.

The container in which the polynucleotide including a sequence encodinga cellular immune response element or fragment thereof may include apackage that is labeled, and the label may bear a notice in the formprescribed by a governmental agency, for example the Food and DrugAdministration, which notice is reflective of approval by the agencyunder Federal law, of the manufacture, use, or sale of thepolynucleotide material therein for human administration.

Federal law requires that the use of pharmaceutical compositions in thetherapy of humans be approved by an agency of the Federal government. Inthe United States, enforcement is the responsibility of the Food andDrug Administration, which issues appropriate regulations for securingsuch approval, detailed in 21 U.S.C. §301-392. Regulation for biologicmaterial, including products made from the tissues of animals isprovided under 42 U.S.C. §262. Similar approval is required by mostforeign countries. Regulations vary from country to country, butindividual procedures are well known to those in the art and thecompositions and methods provided herein preferably comply accordingly.

The dosage to be administered depends to a large extent on the conditionand size of the subject being treated as well as the frequency oftreatment and the route of administration. Regimens for continuingtherapy, including dose and frequency may be guided by the initialresponse and clinical judgment. The parenteral route of injection intothe interstitial space of tissues is preferred, although otherparenteral routes, such as inhalation of an aerosol formulation, may berequired in specific administration, as for example to the mucousmembranes of the nose, throat, bronchial tissues or lungs.

As such, provided herein is a pharmaceutical product which may include apolynucleotide including a sequence encoding a cellular immune responseelement or fragment thereof in solution in a pharmaceutically acceptableinjectable carrier and suitable for introduction interstitially into atissue to cause cells of the tissue to express a cellular immuneresponse element or fragment thereof, a container enclosing thesolution, and a notice associated with the container in form prescribedby a governmental agency regulating the manufacture, use, or sale ofpharmaceuticals, which notice is reflective of approval by the agency ofmanufacture, use, or sale of the solution of polynucleotide for humanadministration.

Indications

The nucleic acid molecules disclosed herein can be used to treatdiseases, conditions or disorders associated with hsp47, such as liverfibrosis, cirrhosis, pulmonary fibrosis, kidney fibrosis, peritonealfibrosis, chronic hepatic damage, and fibrillogenesis and any otherdisease or conditions that are related to or will respond to the levelsof hsp47 in a cell or tissue, alone or in combination with othertherapies. As such, compositions, kits and methods disclosed herein mayinclude packaging a nucleic acid molecule disclosed herein that includesa label or package insert. The label may include indications for use ofthe nucleic acid molecules such as use for treatment or prevention ofliver fibrosis, peritoneal fibrosis, kidney fibrosis and pulmonaryfibrosis, and any other disease or conditions that are related to orwill respond to the levels of hsp47 in a cell or tissue, alone or incombination with other therapies. A label may include an indication foruse in reducing expression of hsp47. A “package insert” is used to referto instructions customarily included in commercial packages oftherapeutic products, that contain information about the indications,usage, dosage, administration, contraindications, other therapeuticproducts to be combined with the packaged product, and/or warningsconcerning the use of such therapeutic products, etc.

Those skilled in the art will recognize that other anti-fibrosistreatments, drugs and therapies known in the art can be readily combinedwith the nucleic acid molecules herein (e.g. siNA molecules) and arehence contemplated herein.

The methods and compositions provided herein will now be described ingreater detail by reference to the following non-limiting examples.

Example 1 Selecting hsp47 Nucleic Acid Molecule Sequences

Nucleic acid molecules (e.g., siNA ≦25 nucleotides) against Hsp47 weredesigned using several computer programs including si RNA at Whitehead(Whitehead Institute for Biomedical Research), IDT siRNA Design(Integrated DNA Technologies), BLOCK-iT RNAi Designer (Invitrogen),siDESIGN Center (Dharmacon), and BIOPREDsi (Friedrich Miescher Institutefor Biomedical Research, part of the Novartis Research Foundation,available at http://www.biopredsi.orgistart.html). The sequences of topscored siRNAs from these programs were compared and selected (seeTable 1) based on the algorithms as well as the sequence homologybetween human and rat. Candidate sequences were validated by in vitroknocking down assays.

Several parameters were considered for selecting a nucleic acid molecule(e.g., a 21-mer siRNA) sequence. Exemplary parameters include:

-   -   1) thermodynamic stability (RISC favors the strand with less        stable 5′ end)    -   2) 30-52% GC content    -   3) positional nucleotide preference:    -   (C/G)NNNNNNNN(A/U)10NNNNNNNN(A/U), where N is any nucleotide    -   4) devoid of putative immunostimulatory motifs    -   5) 2-nucleotide 3′ overhang    -   6) position of siRNA within the transcript (preferably within        cDNA region)    -   7) sequence specificity (checked by using BLAST)    -   8) variations in single nucleotide by checking SNP database

siRNA sequences having ≦25 nucleotides were designed based on theforegoing methods. Corresponding Dicer substrate siRNA (e.g., ≧26nucleotides) were designed based on the smaller sequences and extend thetarget site of the siNA ≦2.5 nucleotide by adding 4 bases to the 3′-endof the sense strand and 6 bases to the 5′-end of the antisense strand.The Dicer substrates that were made generally have a 25 base sensestrand a 27 base antisense strand with an asymmetric blunt ended and3′-overhang molecule. The sequences of the sense and the anti-sensestrand without base modification (base sequence) and with modifications(experimental sequence) are provided in Table 1.

Example 2

In order to screen for the potent of various siNA molecules against boththe human and rat hsp47 genes, various reporter cell lines wereestablished by lenti-viral induction of human HSP47 cDNA-greenfluorescent protein (GFP) or rat GP46 cDNA-GFP construct into 293,HT1080, human HSC line hTERT, or NRK cell lines. These cell lines werefurther evaluated by siRNA against GFP. The remaining fluorescencesignal was measured and normalized to scrambled siRNA (Ambion) andsubsequently normalized to cell viability. The results showed that siRNAagainst GFP knocks down the fluorescence to different extent indifferent cell lines (FIG. 1). 293_HSP47-GFP and 293_GP46-GFP cell lineswere selected for siHsp47 screening due to their ease of transfectionand sensitivity to fluorescence knockdown.

siRNA Transfection:

Cells were transfected with 1.5 pmol per well of siNA against GFP in96-well tissue culture plates using Lipofectamine RNAiMAX (Invitrogen)in a reverse transfection manner. Cells were seeded at 6,000 cells perwell and mixed with the siNA complexs. Fluorescence readings were takenafter 72 hours incubation on a Synergy 2 Multi-Mode Microplate Reader(BioTek).

Cell Viability Assay:

Cells treated with or without siNA were measured for viability after 72hours incubation using CellTiter-Glo Luminescent Cell Viability AssayKit according to the manual (Promega). The readings were normalized tosamples treated with scrambled siNA molecules.

Example 3 Evaluation of Inhibitory Efficiency of siHsp47 on hsp47Expression in Reporter Cell Lines

siNAs against hsp47 were evaluated for their inhibitory efficiency in293 HSP47-GFP and 293_GP46-GFP cell lines by evaluating the change influorescent signal from the reporter GFP. The experiments were carriedout as described in Example 2. The fluorescent signals were normalizedto fluorescent signals from cells treated with scrambled siRNA (Ambion)which served as a control. The results indicate that the tested hsp47siNA molecules were effective in inhibiting hsp47 mRNA in both celllines. However, siNA against GP46 mRNA (as published in the 2008 Sato etal paper) was effective only in the 293 GP46-GFP cell line. The resultsare shown in FIG. 2 A-B.

The 293_HSP47-GFP and 293_GP46-GFP cell lines treated with siRNA againsthsp47 and gp46 were evaluated for viability using the methods describedin Example 2. The cell viability was normalized to cells treated withscrambled siRNA (Ambion). The results indicate that the cell viabilitywas not affected significantly by the treatment with siNA molecules.However, the cell viability of 293_HSP47-GFP cell lines treated withdifferent hsp47 siNA molecules varied depending on the siNA moleculesused, while the viability of 293_GP46-GFP cell lines were similar.Viability for 293 HSP47-GFP cells were lower for siHsp47-6 and Hsp47-7treated cells than the rest. The results are shown in FIG. 2C-D.

Example 4 Evaluation of siHsp47 Inhibitory Effect on hsp47 mRNA byTaqMan® qPCR

In Example 3, the knock down efficiency of siHsp47s in reporter celllines was evaluated by change in fluorescent signal. To validate theresults at the mRNA level, siRNAs targeting endogenous hsp47 weretransfected into cells of the human HSC cell line hTERT usingLipofectamine RNAiMAX (Invitrogen) in a reverse transfection manner asdescribed in Example 2.

The hsp47 mRNA level was evaluated for knock down efficiency of thevarious tested siHsp47 siNA molecules. Briefly, mRNA were isolated fromhTERT cells after 72 hours after transfection using an RNeasy mini kit(Qiagen). The level of hsp47 mRNA was determined by reversetranscription coupled with quantitative PCR using TaqMan® probes.Briefly, cDNA synthesis was carried out using High-Capacity cDNA ReverseTranscription Kit (ABI) according to the manufacturer's instruction, andsubjected to TaqMan Gene Expression Assay (ABI, hsp47 assay IDHs01060395_g1). The level of hsp47 mRNA was normalized to the level ofGAPDH mRNA according to the manufacturer's instruction (ABI). Theresults indicate that siHsp47-C was the most effective among all thehsp47 siRNAs, siHsp47-2 and siHsp47-2d were the next most effective. Thecombinations of siHsp47-1 with siHsp47-2 or siHsp47-1 with siHsp47-2dwere more effective than siHsp47-1 alone. The results are shown in FIG.3.

Example 5 Validation of siHsp47 Knock Down Effect at the Protein Level

The inhibitory effect of different Hsp47 siNA molecules (siHsp47) onhsp47 mRNA expression were validated at the protein level by measuringthe HSP47 in hTERT cells transfected with different siHsp47.Transfection of hTERT cells with different siHsp47 were performed asdescribed in Example 2. Transfected hTERT cells were lysed and the celllysate were clarified by centrifugation. Proteins in the clarified celllysate were resolved by SDS polyacrylamide gel electrophoresis. Thelevel of HSP47 protein in the cell lysate were determined using ananti-HSP47 antibody (Assay Designs) as the primary antibody, Goatanti-mouse IgG conjugated with HRP (Millipore) as the secondaryantibody, and subsequently detected by Supersignal West PicoChemiluminescence kit (Pierce). Anti-actin antibody (Abeam) was used asa protein loading control. The result showed significant decrease in thelevel of Hsp47 protein in cells treated with siHsp47-C, siHsp47-2d,alone or combination of siHsp47-1 with siHsp47-2d.

Example 6 Downregulation of Collagen I expression by hsp47 siRNA

To determine the effect of siHsp47 on collagen I expression level,collagen I mRNA level in hTERT cells treated with different siRNAagainst hsp47 was measured. Briefly, hTERT cells were transfected withdifferent siHsp47 as described in Example 2. The cells were lysed after72 hours and mRNA were isolated using RNeasy mini kit according to themanual (Qiagen). The level of collagen 1 mRNA was determined by reversetranscription coupled with quantitative PCR using TaqMan® probes.Briefly, cDNA synthesis was carried out using High-Capacity cDNA ReverseTranscription Kit (ABI) according to the manual, and subjected to TaqManGene Expression Assay (ABI, COL1A1 assay ID Hs01076780_g1). The level ofcollagen I mRNA was normalized to the level of GAPDH mRNA according tothe manufacturer's instruction (ABI). The signals were normalized to thesignal obtained from cells transfected with scrambled siNA. The resultindicated that collagen I mRNA level is significantly reduced in thecells treated with some of the candidates siHsp47-2, siHsp47-2d, andtheir combination with siHsp47-1 and shown in FIG. 4.

Example 7 Immunofluorescence Staining of hsp47 siRNA Treated hTERT Cells

To visualize the expression of two fibrosis markers, collagen 1 andalpha-smooth muscle actin (SMA), in hTERT cells transfected with orwithout siHsp47, the cells were stained with rabbit anti-collagen Iantibody (Abeam) and mouse anti-alpha-SMA antibody (Sigma). Alexa Fluor594 goat anti-mouse IgG and Alexa Fluor 488 goat anti-rabbit IgG(Invitrogen (Molecular Probes)) were used as secondary antibodies tovisualize collagen I (green) and alpha-SMA (red). Hoescht was used tovisualize nuleus (blue). The results indicate correlation between siRNAknocking down of some of the target genes and collagen/SMA expression.

Example 8 In Vivo Testing of siHSP47 in Animal Models of Liver Fibrosis

siRNA for Rat Liver Cirrhosis Treatment

The siRNA duplex sequence for HSP47 (siHSP47C) is as listed below.

Sense (5′->3′) ggacaggccucuacaacuaTT Antisense (5′->3′)uaguuguagaggccuguccTT

10 mg/ml siRNA stock solution was prepared by dissolving in nucleasefree water (Ambion). For treatment of cirrhotic rats, siRNA wasformulated with vitamin A-coupled liposome as described by Sato et al(Sato Y. et al. Nature Biotechnology 2008. Vol. 26, p 431) in order totarget activated hepatic stellate cells that produce collagen. Thevitamin A (VA)-liposome-siRNA formulation consists of 0.33 μmol/ml ofVA, 0.33 μmol/ml of liposome (Coatsome EL-01-D, NOF Corporation) and 0.5μg/μl of siRNA in 5% glucose solution.

The liver cirrhosis animal model was reported by Sato et al (Sato Y. etal. Nature Biotechnology 2008. Vol. 26, p 431). 4 week-old male SD ratswere induced with liver cirrhosis with 0.5% dimethylnitrosoamine (DMN)(Wako Chemicals, Japan) in phosphate-buffered saline (PBS). A dose of 2ml/kg per body weight was administered intraperitoneally for 3consecutive days per week on days 0, 2, 4, 7, 9, 11, 14, 16, 18, 21, 23,25, 28, 30, 32, 34, 36, 38 and 40

siRNA treatment: siRNA treatment was carried out from day 32 and for 5intravenous injections. In detail, rats were treated with siRNA onday-32, 34, 36, 38 and 40. Then rats were sacrificed on day-42 or 43. 3different siRNA doses (1.5 mg siRNA per kg body weight, 2.25 mg siRNAper kg body weight, 3.0 mg siRNA per kg body weight) were tested. Detailof tested groups and number of animals in each group are as follows:

-   1) Cirrhosis was induced by DMN injection, then 5% glucose was    injected instead of siRNA) (n=10)-   2) VA-Lip-siHSP47C1.5 mg/kg (n=10)-   3) VA-Lip-siHSP47C 2.25 mg/kg (n=10)-   4) VA-Lip-siHSP47C 3.0 mg/Kg (n=10)-   5) Sham (PBS was injected instead of DMN. 5% Glucose was injected    instead of siRNA) (n=6)-   6) No treatment control (Intact) (n=6)

VA-Lip Refers to Vitamin A—Liposome Complex.

Evaluation of therapeutic efficacy: On day 43, 2 out of 10 animals inthe “diseased rat” group and 1 out of 10 animals in “VA-Lip-siHSP47CsiRNA 1.5 mg/kg” died due to development of liver cirrhosis. Theremainder of the animals survived. After rats were sacrificed, livertissues were fixed in 10% formalin. Then, the left lobule of each liverwas embedded in paraffin for histology. Tissue slides were stained withSirius red, and hematoxylin and eosin (HE). Sirius red staining wasemployed to visualize collagen-deposits and to determine the level ofcirrhosis. HE staining was for nuclei and cytoplasm as counter-staining.Each slide was observed under microscope (BZ-8000, Keyence Corp. Japan)and percentage of Sirius red-stained area per slide was determined byimage analysis software attached to the microscope. At least 4 slidesper each liver were prepared for image analysis, and whole area of eachslide (slice of liver) was captured by camera and analyzed. Statisticalanalysis was carried out by Student's t-test.

Results: FIG. 5 shows the fibrotic areas. The area of fibrosis in“diseased rats” was higher than in the “sham” or “no treatment control”groups. Therefore, DMN treatment induced collagen deposition in theliver, which was a typical observation of liver fibrosis. However, thearea of fibrosis was significantly reduced by the treatment of siRNAtargeting HSP47 gene, compared with “disease rat” group (FIG. 5). Thisresult indicates that siRNA as disclosed herein has a therapeuticefficacy in actual disease.

Additional siRNA compounds are tested in the liver fibrosis animalmodel, and were shown to reduce the fibrotic area in the liver.

Example 9 Generation of Sequences for Active Double Stranded RNACompounds to HSP47/SERPINH1 and Production of the siRNAs Shown in TablesA-18, A-19, and B-E

Using proprietary algorithms and the known sequence of the target genes,the sequences of many potential siRNAs were generated. The sequencesthat have been generated using this method are complementary to thecorresponding mRNA sequence.

Duplexes are generated by annealing complementary single strandedoligonucleotides. In a laminar flow hood, a ˜500 μM Stock Solution ofsingle stranded oligonucleotide is prepared by diluting in WFI (waterfor injection, Norbrook). Actual ssRNA concentrations are determined bydiluting each 500 μM ssRNA 1:200 using WFI, and measuring the OD usingNano prop. The procedure is repeated 3 times and the averageconcentration is calculated. The Stock Solution was then diluted to afinal concentration of 250 μM. Complementary single strands wereannealed by heating to 850C and allowing to cool to room temperatureover at least 45 minutes. Duplexes were tested for complete annealing bytesting 5 μl on a 20% polyacrylamide gel and staining. Samples werestored at 800C.

Tables A-18, A-19 and B-E provide siRNAs for HSP47/SERPINH1. For eachgene there is a separate list of 19-mer siRNA sequences, which areprioritized based on their score in the proprietary algorithm as thebest sequences for targeting the human gene expression.

The following abbreviations are used in the Tables A-18, A-19 and B-Eherein: “other spec or Sp.” refers to cross species identity with otheranimals: D-dog, Rt-rat, Rb-Rabbit, Rh-rhesus monkey, P-Pig, M-Mouse;ORF: open reading frame. 19-mers, and 18+1-mers refer to oligomers of 19and 18+1 (U in position 1 of Antisense, A in position 19 of sensestrand) ribonucleic acids in length, respectively.

In Vitro Testing of the siRNA Compounds for the Target Genes

Low-Throughput-Screen (LTS).for siRNA Oligos Directed to Human and RatSERPINH1 Gene.

Cell Lines: Human prostate adenocarcinoma PC3 cells (ATCC, Cat#CRL-1435) were grown in RPMI medium supplemented with 10% FBS and 2 mML-Glutamine and human epithelial cervical cancer HeLa cells (ATCC,Cat#CCL-2) were maintained in Dulbecco's modified Eagle's medium (DMEM)supplemented with 10% FBS, 2 mM L-glutamine. Cells were maintained at37° C. in 5% CO₂.

About 2×10⁵ human PC-3 cells endogenously expressing SERPINH1 gene, wereinoculated in 1.5 mL growth medium in order to reach 30-50% confluenceafter 24 hours. Cells were transfected with Lipofectamine™2000 reagentto a final concentration of 0.01-5 nM per transfected cells. Cells wereincubated at 37±1° C., 5% CO₂ for 48 hours. siRNA transfected cells wereharvested and RNA was isolated using EZ-RNA kit [Biological Industries(#20-410-100)].

Reverse transcription was performed as follows: Synthesis of cDNA wasperformed and human SERPINH1 mRNA levels were determined by Real TimeqPCR and normalized to those of the Cyclophilin A (CYNA, PPIA) mRNA foreach sample. siRNA activity was determined based on the ratio of theSERPINH1 mRNA quantity in siRNA-treated samples versus non-transfectedcontrol samples.

The most active sequences were selected from further assays. From TableA-18 siRNA compounds SERPINH1_(—)2, SERPINH1_(—)6, SERPINH1_(—)13,SERPINH1_(—)45 SERPINH1_(—)45a, SERPINH1_(—)51, SERPINH1_(—)51a,SERPINH1_(—)52 and SERPINH1_(—)86 were selected as preferred compounds.From Table A-19 siRNA compounds SERPINH1_(—)4, SERPINH1_(—)12,SERPINH1_(—)18, SERPINH1_(—)30, SERPINH1_(—)58 and SERPINH1_(—)88 wereselected as preferred compounds.

Other preferred compounds include SERPINH1_(—)50, SERPINH1_(—)67,SERPINH1_(—)73, SERPINH1_(—)74.

IC₅₀ Values for the LTS Selected SERPINII1 siRNA Oligos

About 2×10⁵ human PC-3 or 0.9×10⁵ rat REF52 cells endogenouslyexpressing SERPINH1 gene, were inoculated in 1.5 mL growth medium inorder to reach 30-50% confluence. Cells were transfected with SERPINH1double stranded RNA compounds (I.e. SERPINH1_(—)2, 4, 6, 12, 13, 18, 45,51, 58, 88) with Lipofectamine™2000 reagent to reach final transfectionconcentrations ranging between 0.0029-100 nM. As negative control cellsare treated with Lipofectamine™2000 reagent or with Syntheticrandomized-sequence, non-targeting siRNA at final concentrations of20-100 nM. Cy3-labeled siRNA transfected cells were used as positivecontrol for transfection efficiency.

Cells were incubated at 37±1° C., 5% CO₂ for 48 hours. siRNA transfectedcells were harvested and RNA was isolated using EZ-RNA kit [BiologicalIndustries (#20-410-100) Reverse transcription: Synthesis of cDNA isperformed and human SERPINH1 mRNA levels were determined by Real TimeqPCR and normalized to those of the Cyclophilin A (CYNA, PPIA) mRNA foreach sample.

The IC₅₀ value of the tested RNAi activity was determined byconstructing a dose-response curve using the activity results obtainedwith the various final siRNA concentrations. The dose response curve wasconstructed by plotting the relative amount of residual SERPINH1 mRNAversus the logarithm of transfected siRNA concentration. The curve iscalculated by fitting the best sigmoid curve to the measured data. Themethod for the sigmoid fit is also known as a 3-point curve fit.

$Y = {{Bot} + \frac{100 - {Bot}}{1 + 10^{{({{{Log}\; {IC}\; 50} - X})} \times {HillSlope}}}}$

where Y is the residual SERPINH1 mRNA response, X is the logarithm oftransfected siRNA concentration, Bot is the Y value at the bottomplateau, LogIC50 is the X value when Y is halfway between bottom and topplateaus and HillSlope is the steepness of the curve.

The percent of inhibition of gene expression using specific siRNAs wasdetermined using qPCR analysis of target gene in cells expressing theendogenous gene. Other siRNA compounds according to Tables A-18, A-19and B-E are tested in vitro where it is shown that these siRNA compoundsinhibit gene expression. Activity is shown as percent residual mRNA;accordingly, a lower value reflects better activity.

In order to test the stability of the siRNA compounds in serum, specificsiRNA molecules were incubated in four different batches of human serum(100% concentration) at 37° C. for up to 24 hours. Samples are collectedat 0.5, 1, 3, 6, 8, 10, 16 and 24 hours. The migration patterns as anindication of were determined at each collection time by polyacrylamidegel electrophoresis (PAGE).

Table 3 shows 1050 (or activity where 1050 not calculated) in human cellline for unmodified double stranded nucleic acid compounds (sense andantisense strand unmodified, dTdT 3′ terminal overhangs) selected fromTables A-18 and A-19.

TABLE 3 AntiSense struc- 0.1 0.5 siRNA 1^(st) position ture IC50 nM nM 5nM SERPINH1_6_S709 U A2 0.019 SERPINH1_12_S709 A A1 0.065SERPINH1_23_S709 U A2 0.377 SERPINH1_54_S709 A A1 0.522 SERPINH1_37_S709U A2 0.11 SERPINH1_73_S709 A A1 0.189 SERPINH1_24_S709 U A2 0.271SERPINH1_55_S709 A A1 0.268 SERPINH1_60_S709 U A2 0.163 SERPINH1_88_S709A A1 0.135 SERPINH1_11_S709 U A2 0.079 SERPINH1_30_S709 A A1 0.093SERPINH1_25_S709 U A2 0.229 SERPINH1_56_S709 A A1 0.469 SERPINH1_5_S709U A2 0.178 SERPINH1_81_S709 G A1 1.404 SERPINH1_52_S709 U A2 0.06SERPINH1_58_S709 A A1 0.304 SERPINH1_2_S709 U A2 0.008 SERPINH1_4_S709 AA1 0.006 SERPINH1_43_S709 U A2 1.403 SERPINH1_67_S709 A A1 2.39SERPINH1_16_S709 U A2 134 95 16 SERPINH1_46_S709 A A1 112 84 28SERPINH1_8_S709 U A2 103 90 39 SERPINH1_85_S709 C A1 166 109 59SERPINH1_45_S709 U A2 0.029 SERPINH1_45a_S1354 A A2 0.051siRNA Knock Down Activity:

About 2×10⁵ human PC-3 cells endogenously expressing SERPINH1 gene wereseeded per well in 6 well plates and allowed to grow for about 24 hr to30-70% confluency. Cells were transfected with the siRNAs being testedat different concentrations using the Lipofectamine™2000 reagent(Invitrogen). The cells were incubated at 37° C. in a 5% CO₂ incubatorfor either 48 h or 72 h. At 48-72 h after transfection cells wereharvested and cell RNA was extracted. Cy3-labeled siRNA duplexes wereused as a positive control for transfection efficiency. Mock cellstreated with Lipofectamine™2000 reagent defined as “Control not activesamples” (negative control) and cells treated with a known active siRNA(HSP47-C) at final concentration of 5 nM defined as “Control activesamples” (positive control). Z′ and controls fold {Fold=mean(Negative)/mean (Positive)} are the means to describe the assayefficiency.

The percent inhibition of target gene expression by each siRNA testedwas determined by Qper analysis of a target mRNA from cells. Reversetranscription was performed by synthesizing cDNA from the cells anddetermining target gene mRNA levels by Real Time qPCR. Measured cellmRNA levels were normalized to those of the Cyclophilin A (CYNA, PPIA)mRNA for each sample. siRNA activity was determined based on the ratioof the target gene mRNA quantity siRNA-treated samples versusnon-transfected control samples. Z′ and controls fold {Fold=mean(Negative)/mean(Positive)} are the means to describe the assayefficiency.

The qPCR results are those that passed QC standards, i.e. the value ofthe standard curve slope was in the interval [−4, −3], R2>0.99, noprimer dimers. Results that did not pass the QC requirements weredisqualified from analysis.

IC50 value of the tested RNAi activity was determined by constructing adose-response curve using the activity results obtained with the variousfinal siRNA concentrations. The dose response curve was constructed byplotting the relative amount of residual SERPINH1 mRNA versus thelogarithm of transfected siRNA concentration, as described above.

On-Target and Off-Target Testing of Double Stranded RNA Molecules:

The psiCHECK system enables evaluation of the guide strand (GS)(antisense) and the passenger strand (PS) (sense strand) to elicittargeted (on-target) and off-targeted effects, by monitoring the changesin expression levels of their target sequences. Four psiCHECK™-2-based(Promega) constructs were prepared for the evaluation of target activityand potential off-target activity of each test molecule GS and PSstrands. In each of the constructs one copy or three copies of eitherthe full target or the seed-target sequence, of test molecule PS or GS,was cloned into the multiple cloning site located downstream of theRenilla luciferase translational stop codon in the 3′-UTR region.

The resulting vectors were termed:

1—GS-CM (guide strand, complete-match) vector containing one copy of thefull target sequence (nucleotide sequence fully complementary to thewhole 19-base sequence of the GS of the test molecule);

2—PS-CM (passenger strand, complete-match) vector containing one copy ofthe full target sequence (nucleotide sequence fully complementary to thewhole 19-base sequence of the PS of the test molecule);

3—GS-SM (guide strand, seed-match) vector containing one copy or threecopies of the seed region target sequence (sequence complementary tonucleotides 1-8 of the GS of the test molecule);

4—PS-SM (passenger strand, seed-match) vector containing one copy of theseed region target sequence (sequence complementary to nucleotides 1-8of the PS of the test molecule).

Nomenclature:

guide strand: strand of siRNA that enters the RISC complex and guidescleavage/silencing of the complementary RNA sequence

seed sequence: Nucleotides 2-8 from the 5′ end of the guide strand.

cm (complete match): DNA fragment fully complementary to the guidestrand of siRNA. This DNA fragment is cloned in 3′UTR of a reporter geneand serves as a target for the straightforward RNA silencing.

sm (seed match): 19-mer DNA fragment with nucleotides ns 12-18 fullycomplementary to the ns 2-8 of the guide strand of siRNA. This DNAfragment is cloned in 3′UTR of a reporter gene and serves as a targetfor the “off-target” silencing.

X1: A single copy of cm or sm cloned in 3′UTR of a reporter gene.

X3 Three copies of cm or sm cloned in 3′UTR of a reporter gene,separated with 4 nucleotides one from another.

TABLE 4 non-limiting examples of psiCHECK cloning targets NomenclatureDescription structure S2a_cm_X1 SERPINH1_2, antisenseCTCGAGGAGACACATGGGTGCTATAG clone name complete match = fullyCGGCCGC SEQ_ID_NO: 2724 complimentary to the XhoI SERPINH1_2 sensestrand NotI SERPINH1_2 antisense strand, a single copy. S2acmS_X1 Thesense strand = the strand of 5′- the S2a_cm_X1 clone to beTCGAGGAGACACATGGGTGCTATAGC expressed in the vector, with SEQ_ID_NO: 2725XhoI and NotI sticky ends. S2acmA_X1 The complimentary (antisense) 5′-strand of the S2a_cm_X1 GGCCGCTATAGCACCCATGTGTCTCC clone, with XhoI andNotI SEQ_ID_NO: 2726 sticky ends. S2a_sm_X1 SERPINH1_2, antisense seedCTCGAGTCTCAAACGTTGTGCTATCG clone name match, a single copy,CGGCCGC SEQ_ID_NO: 2727 nucleotides 12-18 are AS(3′-CTCTGTGTACCCACGATAT)complimentary to the SEQ_ID_NO: 2728 nucleotides 2-8 of the seedSERPINH1_2 antisense strand. S2s_cm_X1 SERPINH1_2, sense completeCTCGAGTATAGCACCCATGTGTCTCG clone name match = fully complimentary toCGGCCGC SEQ_ID_NO: 2729 the SERPINH1_2 sense strand ≡ XhoI SERPINH1_2antisense strand NotI antisense strand, a single copy. S2s_sm_X1SERPINH1_2, sense seed CTCGAGGCGATACAAACTGTGTCTAG clone name match, asingle copy, CGGCCGC SEQ_ID_NO: 2730 nucleotides 12-18 areS(3′-ATATCGTGGGTACACAGAG) complimentary to the SEQ_ID_NO: 2731nucleotides 2-8 of the seed SERPINH1_2 sense strand. S2a_sm_X3SERPINH1_2, antisense seed CTCGAGTCTCAAACGTTGTGCTATCttc clone namematch, a triple copy cTCTCAAACGTTGTGCTATCttccTCTCAAACGTTGTGCTATCGCGGCCGC SEQ_ID_NO: 2732 (ttcc - a spacer) Sts_sm_X3SERPINH1_2, sense seed CTCGAGGCGATACAAACTGTGTCTAtt clone name match, atriple copy ccGCGATACAAACTGTGTCTAttccGCG ATACAAACTGTGTCTAGCGGCCGCSEQ_ID_NO: 2733 (ttcc - a spacer)

The target sequences are cloned using the XhoI and NotI compatiblerestriction enzyme sites. Annealing mixtures are prepared in tightlyclosed 0.5 ml Eppendorf tubes, heated in a water bath to 850C, submergedinto the boiling water bath and finally gradually cooled to roomtemperature.

Ligation: The double stranded oligonucleotide generated by the annealingprocedure is ligated to the linearized (by XhoI and NotI) psiCHECK™-2,and transfected into cells using standard techniques. Positive colonieswere identified and sequenced for verification of insert sequence. Table5 shows nucleotide sequences of inserted oligonucleotides.

TABLE 5 Clone Full SEQ ID siRNA name NO: Oligonucleotide sequence(5′>3′) SERPINH1_11 S11s_cm_X1 2734 GGCCGCCGGACAGGCCTCTACAACAC 2735TCGAGTGTTGTAGAGGCCTGTCCGGC S11a_cm_X1 2736 GGCCGCTGTTGTAGAGGCCTGTCCGC2737 TCGAGCGGACAGGCCTCTACAACAGC S11s_sm_X1 2738GGCCGCAGGACAGGAAGAGCACCACC 2739 TCGAGGTGGTGCTCTTCCTGTCCTGC S11a_sm_X12740 GGCCGCGGTTGTAGCTTAAGGGAATC 2741 TCGAGATTCCCTTAAGCTACAACCGCS11s_sm_X3 2742 GGCCGCAGGACAGGAAGAGCACCACGGAAAGGACAGGAAGAGCACCACGGAAAGGACAGGAAGAGCACCACC 2743TCGAGGTGGTGCTCTTCCTGTCCTTTCCGTGGTGCTCTTCCTGTCCTTTCCGTGGTGCTCTTCCTGTCCTGC S11a_sm_X3 2744GGCCGCGGTTGTAGCTTAAGGGAATGGAAGGTTGTAGCTTAAGGGAATGGAAGGTTGTAGCTTAAGGGAATC 2745TCGAGATTCCCTTAAGCTACAACCTTCCATTCCCTTAAGCTACAACCTTCCATTCCCTTAAGCTACAACCGC SERPINH1_30 S30s_cm_X1 2746GGCCGCCGGACAGGCCTCTACAACTC 2747 TCGAGAGTTGTAGAGGCCTGTCCGGC S30a_cm_X12748 GGCCGCAGTTGTAGAGGCCTGTCCGC 2749 TCGAGCGGACAGGCCTCTACAACTGCSERPINH1_2 S2s_cm_X1 2750 GGCCGCGAGACACATGGGTGCTATAC 2751TCGAGTATAGCACCCATGTGTCTCGC S2a_cm_X1 2752 GGCCGCTATAGCACCCATGTGTCTCC2753 TCGAGGAGACACATGGGTGCTATAGC S2s_sm_X1 2754GGCCGCTAGACACAGTTTGTATCGCC 2755 TCGAGGCGATACAAACTGTGTCTAGC S2a_sm_X12756 GGCCGCGATAGCACAACGTTTGAGAC 2757 TCGAGTCTCAAACGTTGTGCTATCGCS2s_sm_X3 2758 GGCCGCTAGACACAGTTTGTATCGCGGAATAGACACAGTTTGTATCGCGGAATAGACACAGTTTGTATCGCC 2759TCGAGGCGATACAAACTGTGTCTATTCCGCGATACAAACTGTGTCTATTCCGCGATACAAACTGTGTCTAGC S2a_sm_X3 2760GGCCGCGATAGCACAACGTTTGAGAGGAAGATAGCACAACGTTTGAGAGGAAGATAGCACAACGTTTGAGAC 2761TCGAGTCTCAAACGTTGTGCTATCTTCCTCTCAAACGTTGTGCTATCTTCCTCTCAAACGTTGTGCTATCGC SERPINH1_4 S4s_cm_X1 2762GGCCGCGAGACACATGGGTGCTATTC 2763 TCGAGAATAGCACCCATGTGTCTCGC S4a_cm_X12764 GGCCGCAATAGCACCCATGTGTCTCC 2765 TCGAGGAGACACATGGGTGCTATTGCSERPINH1_6 S6s_cm X1 2766 GGCCGCACAAGATGCGAGACGAGTAC 2767TCGAGTACTCGTCTCGCATCTTGTGC S6a_cm_X1 2768 GGCCGCTACTCGTCTCGCATCTTGTC2769 TCGAGACAAGATGCGAGACGAGTAGC S6s_sm_X1 2770GGCCGCCCAAGATGATCTAATCTGCC 2771 TCGAGGCAGATTAGATCATCTTGGGC S6a_sm_X12772 GGCCGCGACTCGTCGATACTAGGTGC 2773 TCGAGCACCTAGTATCGACGAGTCGCS6s_sm_X3 2774 TCGAGGCAGATTAGATCATCTTGGTTCCGCAGATTAGATCATCTTGGTTCCGCAGATTAGATCATCTTGGGC 2775GGCCGCCCAAGATGATCTAATCTGCGGAACCAAGATGATCTAATCTGCGGAACCAAGATGATCTAATCTGCC S6a_sm_X3 2776GGCCGCGACTCGTCGATACTAGGTGGGAAGACTCGTCGATACTAGGTGGGAAGACTCGTCGATACTAGGTGC 2777TCGAGCACCTAGTATCGACGAGTCTTCCCACCTAGTATCGACGAGTCTTCCCACCTAGTATCGACGAGTCGC SERPINH1_12 S12s_cm_X1 2778GGCCGCACAAGATGCGAGACGAGTTC 2779 TCGAGAACTCGTCTCGCATCTTGTGC S12a_cm_X12780 GGCCGCAACTCGTCTCGCATCTTGTC 2781 TCGAGACAAGATGCGAGACGAGTTGCSERPINH1_45a S450s_cmX1 2782 GGCCGCACTCCAAGATCAACTTCCTC 2783TCGAGAGGAAGTTGATCTTGGAGTGC (SERPINH1_45_S450) S450a_cmX1 2784GGCCGCAGGAAGTTGATCTTGGAGTC 2785 TCGAGACTCCAAGATCAACTTCCTGC S450s_smX12786 GGCCGCCCTCCAAGCGACCATGAAGC 2787 TCGAGCTTCATGGTCGCTTGGAGGGCS450a_smX1 2788 GGCCGCCGGAAGTTTCGATGTTCTGC 2789TCGAGCAGAACATCGAAACTTCCGGC S450s_smX3 2790GGCCGCCCTCCAAGCGACCATGAAGGGAACCTCCAAGCGACCATGAAGGGAACCTCCAAGCGACCATGAAGC 2791TCGAGCTTCATGGTCGCTTGGAGGTTCCCTTCATGGTCGCTTGGAGGTTCCCTTCATGGTCGCTTGGAGGGC S450a_smX3 2792GGCCGCCGGAAGTTTCGATGTTCTGGGAACGGAAGTTTCGATGTTCTGGGAACGGAAGTTTCGATGTTCTGC 2793TCGAGCAGAACATCGAAACTTCCGTTCCCAGAACATCGAAACTTCCGTTCCCAGAACATCGAAACTTCCGGC SERPINH1_51 S51s_cm_X1 2794GGCCGCTCCTGAGACACATGGGTGAC 2795 TCGAGTCACCCATGTGTCTCAGGAGC S51a_cm_X12796 GGCCGCTCACCCATGTGTCTCAGGAC 2797 TCGAGTCCTGAGACACATGGGTGAGCS51s_sm_X1 2798 GGCCGCGCCTGAGAACACGTGTGTCC 2799TCGAGGACACACGTGTTCTCAGGCGC S51a_sm_X1 2800 GGCCGCGCACCCATTGTGATACTTCC2801 TCGAGGAAGTATCACAATGGGTGCGC S51s_sm_X3 2802GGCCGCGCCTGAGAACACGTGTGTCGGAAGCCTGAGAACACGTGTGTCGGAAGCCTGAGAACACGTGTGTCC 2803TCGAGGACACACGTGTTCTCAGGCTTCCGACACACGTGTTCTCAGGCTTCCGACACACGTGTTCTCAGGCGC S51a_sm_X3 2804GGCCGCGCACCCATTGTGATACTTCGGAAGCACCCATTGTGATACTTCGGAAGCACCCATTGTGATACTTCC 2805TCGAGGAAGTATCACAATGGGTGCTTCCGAAGTATCACAATGGGTGCTTCCGAAGTATCACAATGGGTGCGC SERPINH1_86 S86s_cm_X1 2806GGCCGCACAGGCCTCTACAACTACAC 2807 TCGAGTGTAGTTGTAGAGGCCTGTGC S86a_cm_X12808 GGCCGCTGTAGTTGTAGAGGCCTGTC 2809 TCGAGACAGGCCTCTACAACTACAGCS86s_sm_X1 2810 GGCCGCACAGGCCTAGCACAAGCACC 2811TCGAGGTGCTTGTGCTAGGCCTGTGC S86a_sm_X1 2812 GGCCGCGGTAGTTGGCTCTGAAGTGC2813 TCGAGCACTTCAGAGCCAACTACCGC S86s_sm_X3 2814GGCCGCACAGGCCTAGCACAAGCACGGAAACAGGCCTAGCACAAGCACGGAAACAGGCCTAGCACAAGCACC 2815TCGAGGTGCTTGTGCTAGGCCTGTTTCCGTGCTTGTGCTAGGCCTGTTTCCGTGCTTGTGCTAGGCCTGTGC S86a_sm_X3 2816GGCCGCGGTAGTTGGCTCTGAAGTGGGAAGGTAGTTGGCTCTGAAGTGGGAAGGTAGTTGGCTCTGAAGTGC 2817TCGAGCACTTCAGAGCCAACTACCTTCCCACTTCAGAGCCAACTACCTTCCCACTTCAGAGCCAACTACCGC SERPINH1_52 S52s_cm_X1 2818GGCCGCGACAAGATGCGAGACGAGAC 2819 TCGAGTCTCGTCTCGCATCTTGTCGC S52a_cm_X12820 GGCCGCTCTCGTCTCGCATCTTGTCC 2821 TCGAGGACAAGATGCGAGACGAGAGCS52s_sm_X1 2822 GGCCGCTACAAGATTATCTCATCTCC 2823TCGAGGAGATGAGATAATCTTGTAGC S52a_sm_X1 2824 GGCCGCGCTCGTCTATACTAGGTGAC2825 TCGAGTCACCTAGTATAGACGAGCGC S52s_sm_X3 2826GGCCGCTACAAGATTATCTCATCTCGGAATACAAGATTATCTCATCTCGGAATACAAGATTATCTCATCTCC 2827TCGAGGAGATGAGATAATCTTGTATTCCGAGATGAGATAATCTTGTATTCCGAGATGAGATAATCTTGTAGC S52a_sm_X3 2828GGCCGCGCTCGTCTATACTAGGTGAGGAAGCTCGTCTATACTAGGTGAGGAAGCTCGTCTATACTAGGTGAC 2829TCGAGTCACCTAGTATAGACGAGCTTCCTCACCTAGTATAGACGAGCTTCCTCACCTAGTATAGACGAGCGC SERPINH1_58 S58s_cm_X1 2830GGCCGCGACAAGATGCGAGACGAGTC 2831 TCGAGACTCGTCTCGCATCTTGTCGC S58a_cm_X12832 GGCCGCACTCGTCTCGCATCTTGTCC 2833 TCGAGGACAAGATGCGAGACGAGTGCSERPINH1_95 S95s_cm_X1 2834 GGCCGCACTCCAAGATCAACTTCCGC 2835TCGAGCGGAAGTTGATCTTGGAGTGC S95a_cm_X1 2836 GGCCGCCGGAAGTTGATCTTGGAGTC2837 TCGAGACTCCAAGATCAACTTCCGGC SERPINH1_96 S96s_cm_X1 2838GGCCGCTCCTGAGACACATGGGTGCC 2839 TCGAGGCACCCATGTGTCTCAGGAGC S96a_cm_X12840 GGCCGCGCACCCATGTGTCTCAGGAC 2841 TCGAGTCCTGAGACACATGGGTGCGCSERPINH1_97 S97s_cm_X1 2842 GGCCGCACAGGCCTCTACAACTACTC 2843TCGAGAGTAGTTGTAGAGGCCTGTGC S97a_cm_X1 2844 GGCCGCAGTAGTTGTAGAGGCCTGTC2845 TCGAGACAGGCCTCTACAACTACTGC

Relevant strands, as described above, were cloned in the 3′UTR of thereporter mRNA, Renilla Luciferase in the psiCHECK™-2 (Promega) vector.XhoI and NotI were used as cloning sites using standard molecularbiology techniques. Each strand was chemically synthesized and annealedby heating to 100° C. and cooled to room temperature. Ligation wascarried out for 3 hours using standard molecular biology techniques andtransformed into E. coli DH5a cells. Resulting colonies were screenedfor presence of plasmid constructs by colony-PCR using relevant primers.Each of the plasmids (vectors) was purified from one positive colony andits sequence was verified.

About 1.3×10⁶ human HeLa cells were inoculated in 10 cm dish. Cells werethen incubated in 37±1° C., 5% CO2 incubator for 24 hours. Growth mediumwas replaced one day post inoculation by 8 mL fresh growth medium andeach plate was transfected with one of the plasmids mentioned above,using Lipofectamine™2000 reagent according manufacturing protocol andincubated for 5 hours at 37±10C and 5% CO2. Following incubation, cellswere re-plated in a 96-well plate at final concentration of 5×103 cellsper well in 80 μL growth medium. 16 hours later, cells were transfectedwith SERPINH1 siRNA molecules using Lipofectamine™2000 reagent atdifferent concentrations ranging from 0.001 nM to 5 nM in a 1000. finalvolume. Mock cells treated with Lipofectamine™2000 reagent with thecorresponding psiCHECK™-2 plasmid defined as “Control not activesamples” (negative control) and cells treated with a known active siRNA(HSP47-C) at final concentration of 5 nM defined as “Control activesamples” (positive control). Z′ and controls fold {Fold=mean(Negative)/mean(Positive)} are the means to describe the assayefficiency.

Cells were then incubated for 48 hours at 37±1° C. and Renilla andFireFly Luciferase activities were measured in each of the siRNAtransfected samples, using Dual-Luciferase® Assay kit (Promega,Cat#E1960) according to manufacturer procedure. The activity of asynthetic siRNA toward this target sequence results either in cleavageand subsequent degradation of the fused mRNA or in translationinhibition of the encoded protein. Measuring the decrease in Renillaluciferase activity thus provides a convenient way of monitoring siRNAeffect while Firefly luciferase, allows normalization of Renillaluciferase expression. Renilla Luciferase activity value was divided byFirefly Luciferase activity value for each sample (normalization).Renilla luciferase activity is finally expressed as the percentage ofthe normalized activity value in tested sample relative to “Control notactive samples”.

Results of TNFα and IL-6 levels in peripheral blood mononuclear cells(PMNC) exposed to unmodified or modified siRNA/Lipofectamine™2000.Results are provided in pg/ml values calculated based on standard curve.“Control Lipofec2000” relates to level of cytokine secretion induced bythe transfection reagent, Lipofectamine™2000. None of the modifiedcompounds levels of cytokines TNFα or IL6 above those of the controltransfection reagent.

Donor II TNFa IL-6 control 162 +/− 280 Control Lipofec2000 308 +/− 75 1303 +/− 440  dsRNA SEQ ID 860 nM 610 2915 NOS: 101 and 168) 287 nM 69634021 unmodified  96 nM 641 2278  32 nM 1095 4126 Compound_4 860 nM 660+/− 227 1166 +/− 280  287 nM 484 +/− 84  1844 +/− 1072  96 nM 571 +/−170 2015 +/− 1667  32 nM 865 +/− 90  2201 +/− 952  Donor I Donor II TNFaIL-6 TNFa IL-6 control 115 +/− 64 162 +/− 280 control Lipofec2000 427+/− 87 1848 +/− 194 308 +/− 75  1303 +/− 440 dsRNA SEQ ID 860 nM 3261014 873 4015 NOS: 60 and 127) 287 nM 305 638 909 3046 unmodified  96 nM546 1007 690 2451  32 nM 707 1331 637 2159 Compound_1 860 nM 491 14801017 4492 287 nM 363 956 981 3126  96 nM 294 840 952 2491  32 nM 355 848902 2779 Donor I TNFa IL-6 control 115 +/− 64 control Lipofec2000 427+/− 87 1848 +/− 194 dsRNA SEQ ID 860 nM 228 553 NOS: 63 and 130) 287 nM395 569 unmodified  96 nM 561 966  32 nM 737 1021 Compound_2 860 nM 5981560 287 nM 621 1440  96 nM 570 1825  32 nM 517 1510 Donor I Donor IITNFa IL-6 TNFa IL-6 control 115 +/− 64 162 +/− 280 control Lipofec2000427 +/− 87 1848 +/− 194 308 +/− 75  1303 +/− 440 dsRNA SEQ ID 860 nM 137225 521 4223 NOS: 98 and 165) 287 nM 750 105 463 3755 unmodified  96 nM504 180 627 2784  32 nM 312 442 711 3084 Compound_3 860 nM 540 2170 14743896 287 nM 698 2428 1000 1864  96 nM 582 1876 1089 1760  32 nM 614 1341724 1044 Ctrl cells 115 +/− 64 162 +/− 280 CL075 (ug/ml) 2 13878 264640.67 8115 28471 17013 0.22 1575 10873 7589 22111 0.074 219 906 1389 7072Results are pg/ml

Data for induction of interferon (IFN) responsive genes, MX1 and IFIT1,unmodified and modified double stranded nucleic acid compounds. Resultsshown are residual (fold of Ctrl Lipofectamine-2000 treated cells) humanIFIT1 and MX1 genes as tested in human PMNC. Data shows that allmodified compounds induced negligible levels of IFN downstream genes,compared to unmodified (_S709) compounds.

Donor II IFIT1 MX1 Ctrl Lipo2000 1 1 dsRNA SEQ ID NOS: 101 and 168)unmodified  32 nM 5.5 3.3  96 nM 7.5 4.3 297 nM 3.9 3.8 860 nM 0.8 0.8Compound_4  32 nM 1.2 +/− 0.5 1.7 +/− 0.35  96 nM 1.1 +/− 0.3 1.5 +/−0.06 297 nM 0.7 +/− 0.3 0.9 +/− 0.7  860 nM 0.6 +/− 0.1 0.9 +/− 0.5 Donor I Donor II IFIT1 MX1 IFIT1 MX1 Ctrl Lipo2000 1 1 1 1 dsRNA SEQ IDNOS: 60 and 127) unmodified  32 nM 27.9 — 2.2 2.7  96 nM 42.1 18.3 4.05.0 297 nM 53.8 18.0 3.4 2.8 860 nM 39.4 16.3 3.3 3.6 Compound_1  32 nM1.2 0.2 0.8 1.3  96 nM 1.3 0.8 1.5 1.1 297 nM 1.1 0.3 1.2 1.6 860 nM 1.00.3 0.3 0.3 Ctrl Lipo2000 1 1.00 1 1 dsRNA SEQ ID NOS: 98 and 165)unmodified  32 nM 29.7 18.5 4.3 4.1  96 nM 39.1 19.2 3.5 297 nM 25.1 9.34.8 5.2 860 nM 3.8 3.7 Compound_3  32 nM 1.4 0.4 1.0 1.4  96 nM 1.7 1.31.3 1.1 297 nM 1.9 1.4 1.1 1.4 860 nM 5.2 2.5 1.1 1.4 Donor I IFIT1 MX1Ctrl Lipo2000 1 1 dsRNA SEQ ID NOS: 63 and 130) unmodified  32 nM 29.617.8  96 nM 31.5 16.1 297 nM 860 nM 36.6 11.4 Compound_2  32 nM 1.6 0.7 96 nM 1.1 1.0 297 nM 2.1 0.2 860 nM 1.8 1.4 Donor I Donor II IFIT1 MX1IFIT1 MX1 Ctrl cells 1 1 1 1 0.125 18 5.4 3.7 0.56 26 11 4.9 4.7 1.7 4114 4.5 5.1 5 24 7 0.9 0.8 0.075 4 2 1.8 1.8 0.12 27 10 4.5 3.6 0.67 214.6 4.2 2 26 12 4.1 3.7

The tables below show activity of Compound_(—)1, Compound_(—)2, Compound3 and Compound_(—)4 compared to unmodified LS709) compounds in ratcells. Results are shown as residual target (% of controlLipofectamine™2000 treated cells) rat SERPINH1 gene in REF52 cells.Results of two separate experiments are shown. Knockdown to target genein rat cells is relevant to testing compounds in animal models of humandisease.

Study_1 Study_2 Ctrl Lipo2000 100 100 dsRNA SEQ ID NOS: 60 0.8 nM  52 36and 127) unmodified  2 nM 25 31 10 nM 16 28 50 nM 8 4 Compound_1 0.8 nM 53 14  2 nM 39 14 10 nM 19 24 50 nM 7 4 dsRNA SEQ ID NOS: 63 0.8 nM  4515 and 130) unmodified  2 nM 28 18 10 nM 13 12 50 nM 12 8 Compound_2 0.8nM  76 78  2 nM 61 68 10 nM 37 28 50 nM 4 dsRNA SEQ ID NOS: 98 0.8 nM 72 65 and 165) unmodified  2 nM 43 41 10 nM 32 42 50 nM 28 27 Compound_30.8 nM  88 30  2 nM 39 24 10 nM 24 23 50 nM 6 23 Study_3 Study_4 CtrlLipo2000 100 100 Compound_4 0.8 nM  66 106  2 nM 35 32 10 nM 10 12 50 nM6 9

Serum Stability Assay

The modified compounds according to the present invention are tested forduplex stability in human serum or human tissue extract, as follows:

siRNA molecules at final concentration of 7 uM are incubated at 370C in100% human serum (Sigma Cat# H4522). (siRNA stock 100 uM diluted inhuman serum 1:14.29 or human tissue extract from various tissue types).Five ul (5 ul) are added to 15ul 1.5×TBE-loading buffer at differenttime points (for example 0, 30 min, 1 h, 3 h, 6 h, 8 h, 10 h, 16 h and24 h). Samples were immediately frozen in liquid nitrogen and kept at−20° C.

Each sample is loaded onto a non-denaturing 20% acrylamide gel, preparedaccording to methods known in the art. The oligos were visualized withethidium bromide under UV light.

Exonuclease Stability Assay

To study the stabilization effect of 3′ non-nucleotide moieties on anucleic acid molecule the sense strand, the antisense strand and theannealed siRNA duplex are incubated in cytosolic extracts prepared fromdifferent cell types.

Extract: HCT116 cytosolic extract (12 mg/ml).Extract buffer: 25 mM Hepes pH-7.3 at 37° C.; 8 mM MgCl; 150 mM NaClwith 1 mM DTT was added fresh immediately before use.

Method: 3.5 ml of test siRNA (100 mM), were mixed with 46.5 ml contain120 mg of HCT116 cytosolic extract. The 46.5 ml consists of 12 ml ofHCT116 extract, and 34.5 ml of the extract buffer supplemented with DTTand protease inhibitors cocktail/100 (Calbiochem, setIII-539134). Thefinal concentration of the siRNA in the incubation tube is 7 mM. Thesample was incubated at 37° C., and at the indicated time point 5 mlwere moved to fresh tube, mixed withl5 ml of 1×TBE-50% Glycerol loadingbuffer, and snap frozen in Liquid N2. The final concentration of thesiRNA in the loading buffer is 1.75 mM (21 ng siRNA/ml). For Analyses bynative PAGE and EtBr staining 50 ng are loaded per lane. For Northernanalyses 1 ng of tested siRNA was loaded per lane.

Innate Immune Response to SERPINH1 siRNA Molecules:

Fresh human blood (at RT) was mixed at 1:1 ratio with sterile 0.9% NaClat RT, and gently loaded (1:2 ratio) on Ficoll (Lymphoprep, Axis-Shieldcat# 1114547). Samples were centrifuged at RT (22° C., 800g) in aswinging centrifuge for 30 minutes, washed with RPMI1640 medium andcentrifuged (RT, 250 g) for 10 minutes. Cells were counted and seeded atfinal concentration of 1.5×10⁶ cell/ml in growth medium (RPMI1640+10%FBS+2 mM L-glutamine+1% Pen-Strep) and incubated for 1 hours at 37° C.before siRNA treatment.

Cells were then treated with the siRNAs being tested at differentconcentrations using the Lipofectamine™2000 reagent (Invitrogen)according manufacturer's instructions and incubated at 370C in a 5% CO2incubator for 24 hours.

As a positive control for IFN response, cells were treated with eitherpoly(I:C), a synthetic analog of double strand RNA (dsRNA) which is aTLR3 ligand (InvivoGen Cat# tlr1-pic) at final concentrations of0.25-5.0 μg/mL or to Thiazolaquinolone (CLO75), a TLR 7/8 ligand(InvivoGen Cat# tlr1-c75) at final concentrations of 0.075-2 μg/mL. Celltreated with Lipofectamine™2000 reagent were used as negative(reference) control for IFN response.

At about 24 hours following incubation, cells were collected andsupernatant was transferred to new tubes. Samples were frozenimmediately in liquid nitrogen and secretion of IL-6 and TNF-α cytokineswas tested using IL-6, DuoSet ELISA kit (R&D System DY2060), and TNF-α,DuoSet ELISA kit (R&D System DY210), according to manufacturer'sinstructions. RNA was extracted from the cell pellets and mRNA levels ofhuman genes IFIT1 (interferon-induced protein with tetratricopeptiderepeats 1) and MX1 (myxovirus (influenza virus) resistance 1,interferon-inducible protein p78) were measured by qPCR. Measured mRNAquantities were normalized to the mRNA quantity of the reference genepeptidylprolyl isomerase A (cyclophilin A; CycloA). Induction ofIFN-signaling was evaluated by comparing the quantity of mRNA from IFIT1and MX1 genes from treated cells, relative to their quantitiesnon-treated cells. The qPCR results are those that passed QC standards,i.e. the value of the standard curve slope was in the interval [−4, −3],R2>0.99, no primer dimers. Results that did not pass the QC requirementswere disqualified from analysis.

Table 6 shows siSERPINH1 compounds. Activity and stability data for someof the compounds is presented in Table 6. The code for the sense andantisense strand structures is presented in Table 7, infra.

TABLE 6 % % residual residual 1. Stability in 5 nM 25 nM Sense strand5->3 AntiSense strand 5->3

ame plasma (h) 001 002 003 004 006 code code SERPINH1_2_S1356 10 16 10 9zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA; mG; rC; mA; rC; rA;rU; rG; rG; rG; rU; rG; rC2p; mC; rC; mA; rU; mG; rU; mG; rU2p; rA2p;rU2p; rA2p rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1357 zidB; rG; rA;rG; rA; rC; rA; rC; mU; rA; mU; rA; rG; mC; rA; mC; rA; rU; rG; rG; rG;rU; rG; rC2p; mC; mC; rA; mU; rG; mU; rG; rU2p; rA2p; rU2p; rA2p mU; mC;mU; rC; zc3p; zc3p$ SERPINH1_2_S1358 16 52 41 zidB; rG; rA; rG; rA; rC;rA; rC; mU; rA; mU; rA; rG; mC; rA; mC; rA; rU; rG; rG; rG; rU; rG;rC2p; mC; rC; rA; mU; rG; mU; rG; rU2p; rA2p; rU2p; rA2p mU; mC; mU; rC;zc3p; zc3p$ SERPINH1_2_S1359 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA;mU; rA; rG; mC; rA; mC; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; rC; rA;mU; rG; mU; rG; rU; rU2p; rA2p; rU2p; rA2p mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1360 10 47 31 8 20 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA;mU; rA; rG; mC; rA; mC; rA; rU; rG; rG; rG; rU; rG; rC2p; rC; mC; rA;mU; rG; mU; rG; rU; rU2p; rA2p; rU2p; rA2p mC; mU; rC; zc3p; zc3p$SERPINH1_2_S1361 8 31 34 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU;rA; rG; rC2p; rA; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; mC; rC; rA; mU;rG; mU; rG; rU2p; rA2p; rU2p; rA2p rU; mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1362 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA; rG;LdC; rA; mC; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; rC; rA; mU; rG; mU;rG; rU; rU2p; rA2p; rU2p; rA2p mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S136317 10 15 25 zc3p; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA; mG; rC;mA; rC; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; rC; mA; rU; mG; rU; mG;rU2p; rA2p; rU2p; rA2p rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1364zc3p; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA; rG; mC; rA; mC; rA;rU; rG; rG; rG; rU; rG; rC2p; mC; mC; rA; mU; rG; mU; rG; rU2p; rA2p;rU2p; rA2p mU; mC; mU; rC; zc3p; zc3p$ SERPINH1_2_S1365 16 41 52 zc3p;rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA; rG; mC; rA; mC; rA; rU; rG;rG; rG; rU; rG; rC2p; mC; rC; rA; mU; rG; mU; rG; rU2p; rA2p; rU2p; rA2pmU; mC; mU; rC; zc3p; zc3p$ SERPINH1_2_S1366 zc3p; rG; rA; rG; rA; rC;rA; rC; mU; rA; mU; rA; rG; mC; rA; mC; rA; rU; rG; rG; rG; rU; rG;rC2p; mC; rC; rA; mU; rG; mU; rG; rU; rU2p; rA2p; rU2p; rA2p mC; rU; mC;zc3p; zc3p$ SERPINH1_2_S1367 16 51 39 zc3p; rG; rA; rG; rA; rC; rA; rC;mU; rA; mU; rA; rG; mC; rA; mC; rA; rU; rG; rG; rG; rU; rG; rC2p; rC;mC; rA; mU; rG; mU; rG; rU; rU2p; rA2p; rU2p; rA2p mC; mU; rC; zc3p;zc3p$ SERPINH1_2_S1368 zc3p; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA;rG; rC2p; rA; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; mC; rC; rA; mU; rG;mU; rG; rU2p; rA2p; rU2p; rA2p rU; mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1369 zc3p; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA; rG;LdC; rA; mC; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; rC; rA; mU; rG; mU;rG; rU; rU2p; rA2p; rU2p; rA2p mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S137017 15 61 20 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA; mG; rC;mA; rC; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; rC; mA; rU; mG; rU; mG;rU2p; rA2p; rU2p; rA2p; zc3p$ rU; mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1371 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA; rG;mC; rA; mC; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; mC; rA; mU; rG; mU;rG; rU2p; rA2p; rU2p; rA2p; zc3p$ mU; mC; mU; rC; zc3p; zc3p$SERPINH1_2_S1372 16 74 66 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU;rA; rG; mC; rA; mC; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; rC; rA; mU;rG; mU; rG; rU2p; rA2p; rU2p; rA2p; zc3p$ mU; mC; mU; rC; zc3p; zc3p$SERPINH1_2_S1373 8 48 65 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU;rA; rG; mC; rA; mC; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; rC; rA; mU;rG; mU; rG; rU; rU2p; rA2p; rU2p; rA2p; zc3p$ mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1374 16 39 110 6 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA;mU; rA; rG; mC; rA; mC; rA; rU; rG; rG; rG; rU; rG; rC2p; rC; mC; rA;mU; rG; mU; rG; rU; rU2p; rA2p; rU2p; rA2p; zc3p$ mC; mU; rC; zc3p;zc3p$ SERPINH1_2_S1375 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA;rG; rC2p; rA; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; mC; rC; rA; mU; rG;mU; rG; rU2p; rA2p; rU2p; rA2p; zc3p$ rU; mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1376 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA; mU; rA; rG;LdC; rA; mC; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; rC; rA; mU; rG; mU;rG; rU; rU2p; rA2p; rU2p; rA2p; zc3p$ mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1377 3 25 5 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA;mG; rC; mA; rC; mC; rA; rU; rG; rG; rG; mU; rG; mC; rC; mA; rU; mG; rU;mG; mC; mU; rA; LdT; rA$ rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1378zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; rG; mC; rA; mC; mC; rA;rU; rG; rG; rG; mU; rG; mC; mC; rA; mU; rG; mU; rG; mC; mU; rA; LdT; rA$mU; mC; mU; rC; zc3p; zc3p$ SERPINH1_2_S1379 zidB; rG; rA; rG; rA; mC;rA; mU; rA; mU; rA; rG; mC; rA; mC; mC; rA; rU; rG; rG; rG; mU; rG; mC;rC; rA; mU; rG; mU; rG; mC; mU; rA; LdT; rA$ mU; mC; mU; rC; zc3p; zc3p$SERPINH1_2_S1380 8 23 33 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA;rG; mC; rA; mC; mC; rA; rU; rG; rG; rG; mU; rG; mC; rC; rA; mU; rG; mU;rG; rU; mC; mU; rA; LdT; rA$ mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1381 1625 56 12 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; rG; mC; rA; mC;mC; rA; rU; rG; rG; rG; mU; rG; rC; mC; rA; mU; rG; mU; rG; rU; mC; mU;rA; LdT; rA$ mC; mU; rC; zc3p; zc3p$ SERPINH1_2_S1382 8 22 31 11 zidB;rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; rG; rC2p; rA; mC; rA; rU; rG;rG; rG; mU; rG; mC; mC; rC; rA; mU; rG; mU; rG; mC; mU; rA; LdT; rA$ rU;mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1383 zidB; rG; rA; rG; rA; mC; rA;mU; rA; mU; rA; rG; LdC; rA; mC; mC; rA; rU; rG; rG; rG; mU; rG; mC; rC;rA; mU; rG; mU; rG; rU; mC; mU; rA; LdT; rA$ mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1384 16 7 20 7 4 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU;rA; mG; rC; mA; rC; mC; rA; rU; rG; rG; rG; mU; rG; mC; rC; mA; rU; mG;rU; mG; rC; mU; rA; mU; rA; zc3p$ rU; mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1385 16 55 37 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA;rG; mC; rA; mC; mC; rA; rU; rG; rG; rG; mU; rG; mC; mC; rA; mU; rG; mU;rG; rC; mU; rA; mU; rA; zc3p$ mU; mC; mU; rC; zc3p; zc3p$SERPINH1_2_S1386 16 42 45 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA;rG; mC; rA; mC; mC; rA; rU; rG; rG; rG; mU; rG; mC; rC; rA; mU; rG; mU;rG; rC; mU; rA; mU; rA; zc3p$ mU; mC; mU; rC; zc3p; zc3p$SERPINH1_2_S1387 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; rG; mC;rA; mC; mC; rA; rU; rG; rG; rG; mU; rG; mC; rC; rA; mU; rG; mU; rG; rU;rC; mU; rA; mU; rA; zc3p$ mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1388 16 2139 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; rG; mC; rA; mC; mC; rA;rU; rG; rG; rG; mU; rG; rC; mC; rA; mU; rG; mU; rG; rU; rC; mU; rA; mU;rA; zc3p$ mC; mU; rC; zc3p; zc3p$ SERPINH1_2_S1389 16 20 27 zidB; rG;rA; rG; rA; mC; rA; mU; rA; mU; rA; rG; rC2p; rA; mC; rA; rU; rG; rG;rG; mU; rG; mC; mC; rC; rA; mU; rG; mU; rG; rC; mU; rA; mU; rA; zc3p$rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1390 zidB; rG; rA; rG; rA; mC;rA; mU; rA; mU; rA; rG; LdC; rA; mC; mC; rA; rU; rG; rG; rG; mU; rG; mC;rC; rA; mU; rG; mU; rG; rU; rC; mU; rA; mU; rA; zc3p$ mC; rU; mC; zc3p;zc3p$ SERPINH1_2_S1687 zidB; rG; rA; rG; rA; rC; rA; mC; mU; rA; mU; rA;mG; rC; mA; rC; rA; rU; rG; rG; rG; mU; rG; rC; mC; rC; mA; rU; mG; rU;mG; mU; rA; mU; rA; zc3p$ rU; mC; rU; rC; zc3p; zc3p$ SERPINH1_2_S169424 zidB; rG; rA; rG; rA; rC; rA; mC; mU; rA; mU; rA; mG; rC; rA2p; rA;rU2p; rG; rG; rG; mU; rG; rC; mC; rC; mA; rU; mG; rU; mG; rC; mU; rA;mU; rA; zc3p$ rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1700 16 zidB; rG;rA; rG; rA; mC; rA; mU; rA; mU; rA; mG; rC; rA2p; mC; rA; rU; rG; rG;rG; mU; rG; rC; mC; rC; mA; rU; mG; rU; mG; rC; mU; rA; rU2p; rA; zc3p$rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1705 10 zidB; rG; rA; rG; rA;rC; rA; rC; mU; rA; mU; rA; mG; rC; rA2p; rA; rU; rG; rG; rG; rU; rG;rC2p; rC; mC; rC; mA; rU; mG; rU; mG; rU2p; rA2p; rU2p; rA2p rU; mC; rU;mC; zc3p; zc3p$ SERPINH1_2_S1707 10 zidB; rG; rA; rG; rA; rC; rA; rC;mU; rA; mU; rA; mG; rC; rA2p; rA; rU; rG; rG; rG; rU; rG; rC2p; rC; mC;rC; mA; rU; mG; rU; mG; rU2p; rA2p; rU2p; rA2p; zc3p$ rU; mC; rU; mC;zc3p; zc3p$ SERPINH1_2_S1754 24 zidB; rG; rA; rG; rA; rC; rA; rC; mU;rA; mU; rA; mG; rC; rA2p; rA; rU; rG; rG; rG; rU; rG; rC2p; rC; mC; rC;mA; rU; mG; rU; mG; rU2p; rA2p; rU2p; rA2p; zc3p rU; mC; rU; mC; zc3p;zc3p$ SERPINH1_2_S1755 24 zidB; rG; rA; rG; rA; rC; rA; rC; rU2p; rA;mU; rA; mG; rC; rA2p; rA; rU; rG; rG; rG; rU; rG; rC2p; rC; mC; rC; mA;rU; mG; rU; rU2p; rA2p; rU2p; rA2p; zc3p mG; rU; mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1756 zidB; rG; rA; rG; rA; rC; rA; rC; rU2p; rA; mU; rA; mG;rC; rA2p; rA; rU; rG; rG; rG; rU; rG; rC2p; rC; mC; rC; mA; rU; mG; rU;rU2p; rA2p; rU2p; rA2p mG; rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1787zidB; rG; rA; rG; rA; rC; rA; rC; dU; rA; mU; rA; mG; rC; rA2p; rC; rA;rU; rG; rG; rG; rU; rG; rC2p; mC; rC; mA; rU; mG; rU; mG; rU2p; rA2p;rU2p; rA2p; zc3p rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_4_S1391 0 58 zidB;rG; rA; rG; rA; rC; rA; rC; rA; mA; rU; mA; rG; mC; rA; mC; rA; rU; rG;rG; rG; rU; rG; rC2p; rC; rC; mA; rU; mG; rU; mG; rU; rU2p; rA2p; rU2p;rU2p mC; rU; mC; zc3p; zc3p$ SERPINH1_4_S1782 zidB; rG; rA; rG; rA; rC;rA; rC; rA; rA; mU; rA; mG; rC; rA2p; rC; rA; rU; rG; rG; rG; rU; rG;rC2p; mC; rC; mA; rU; mG; rU; mG; rU2p; rA2p; rU2p; rU2p; zc3p rU; mC;rU; mC; zc3p; zc3p$ SERPINH1_6_S1356 zidB; rA; rC; rA; rA; rG; rA; rU;mU; rA; mC; rU; mC; rG; mU; rC; rG; rC; rG; rA; rG; rA; rC; rG2p; mU;rC; mG; rC; mA; rU; mC; rA2p; rG2p; rU2p; rA2p rU; mU; rG; mU; zc3p;zc3p$ SERPINH1_6_S1363 zc3p; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU;mC; rG; mU; rC; rG; rC; rG; rA; rG; rA; rC; rG2p; mU; rC; mG; rC; mA;rU; mC; rA2p; rG2p; rU2p; rA2p rU; mU; rG; mU; zc3p; zc3p$SERPINH1_6_S1370 zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC;rG; mU; rC; rG; rC; rG; rA; rG; rA; rC; rG2p; mU; rC; mG; rC; mA; rU;mC; rA2p; rG2p; rU2p; rA2p; zc3p$ rU; mU; rG; mU; zc3p; zc3p$SERPINH1_6_S1414 zc3p; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; mU; mC;rG; mU; rG; rC; rG; rA; rG; rA; rC; rG2p; mC; mU; mC; rG; mC; rA; mU;rA2p; rG2p; rU2p; rA2p mC; mU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1415zc3p; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; mU; mC; rG; mU; rG; rC;rG; rA; rG; rA; rC; rG2p; mC; mU; rC; rG; mC; rA; mU; mC; rA2p; rG2p;rU2p; rA2p mU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1416 zc3p; rA; rC;rA; rA; rG; rA; rU; mU; rA; mC; mU; mC; rG; mU; rG; rC; rG; rA; rG; rA;rC; rG2p; mC; mU; rC; rG; mC; rA; rU; mC; rA2p; rG2p; rU2p; rA2p mU; mU;rG; rU; zc3p; zc3p$ SERPINH1_6_S1417 zc3p; rA; rC; rA; rA; rG; rA; rU;mU; rA; mC; rU; mC; rG; mU; mC; rG; rC; rG; rA; rG; rA; rC; rG2p; rU;mC; rG; mC; mA; rU; mC; rA2p; rG2p; rU2p; rA2p rU; mU; rG; rU; zc3p;zc3p$ SERPINH1_6_S1418 zc3p; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU;mC; rG; LdT; rC; rG; rC; rG; rA; rG; rA; rC; rG2p; mU; rC; rG; mC; mA;rU; mC; rA2p; rG2p; rU2p; rA2p rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1419 zc3p; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC;rG; rU2p; rG; rC; rG; rA; rG; rA; rC; rG2p; rC; mU; rC; rG; mC; mA; rU;mC; rA2p; rG2p; rU2p; rA2p rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1420zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; mU; mC; rG; mU; rG; rC;rG; rA; rG; rA; rC; rG2p; mC; mU; mC; rG; mC; rA; mU; rA2p; rG2p; rU2p;rA2p mC; mU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1421 zidB; rA; rC; rA;rA; rG; rA; rU; mU; rA; mC; mU; mC; rG; mU; rG; rC; rG; rA; rG; rA; rC;rG2p; mC; mU; rC; rG; mC; rA; mU; mC; rA2p; rG2p; rU2p; rA2p mU; mU; rG;rU; zc3p; zc3p$ SERPINH1_6_S1422 zidB; rA; rC; rA; rA; rG; rA; rU; mU;rA; mC; mU; mC; rG; mU; rG; rC; rG; rA; rG; rA; rC; rG2p; mC; mU; rC;rG; mC; rA; rU; mC; rA2p; rG2p; rU2p; rA2p mU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1423 zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC;rG; mU; mC; rG; rC; rG; rA; rG; rA; rC; rG2p; rU; mC; rG; mC; mA; rU;mC; rA2p; rG2p; rU2p; rA2p rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1424zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC; rG; LdT; rC; rG;rC; rG; rA; rG; rA; rC; rG2p; mU; rC; rG; mC; mA; rU; mC; rA2p; rG2p;rU2p; rA2p rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1425 zidB; rA; rC;rA; rA; rG; rA; rU; mU; rA; mC; rU; mC; rG; rU2p; rG; rC; rG; rA; rG;rA; rC; rG2p; rC; mU; rC; rG; mC; mA; rU; mC; rA2p; rG2p; rU2p; rA2p rU;mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1426 zidB; rA; rC; rA; rA; rG; rA;rU; mU; rA; mC; mU; mC; rG; mU; rG; rC; rG; rA; rG; rA; rC; rG2p; mC;mU; mC; rG; mC; rA; mU; rA2p; rG2p; rU2p; rA2p; zc3p$ mC; mU; mU; rG;rU; zc3p; zc3p$ SERPINH1_6_S1427 zidB; rA; rC; rA; rA; rG; rA; rU; mU;rA; mC; mU; mC; rG; mU; rG; rC; rG; rA; rG; rA; rC; rG2p; mC; mU; rC;rG; mC; rA; mU; mC; rA2p; rG2p; rU2p; rA2p; zc3p$ mU; mU; rG; rU; zc3p;zc3p$ SERPINH1_6_S1428 zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; mU;mC; rG; mU; rG; rC; rG; rA; rG; rA; rC; rG2p; mC; mU; rC; rG; mC; rA;rU; mC; rA2p; rG2p; rU2p; rA2p; zc3p$ mU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1429 zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC;rG; mU; mC; rG; rC; rG; rA; rG; rA; rC; rG2p; rU; mC; rG; mC; mA; rU;mC; rA2p; rG2p; rU2p; rA2p; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1430 zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC;rG; LdT; rC; rG; rC; rG; rA; rG; rA; rC; rG2p; mU; rC; rG; mC; mA; rU;mC; rA2p; rG2p; rU2p; rA2p; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1431 zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC;rG; rU2p; rG; rC; rG; rA; rG; rA; rC; rG2p; rC; mU; rC; rG; mC; mA; rU;mC; rA2p; rG2p; rU2p; rA2p; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1432 0 6 19 15 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU;mC; rG; mU; rC; mU; rG; rC; rG; rA; rG; rA; mC; mU; rC; mG; rC; mA; rU;mC; rG; rA; rG; LdT; rA$ rU; mU; rG; mU; zc3p; zc3p$ SERPINH1_6_S1435 637 46 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; mU; mC; rG; mU; mU; rG;rC; rG; rA; rG; rA; mC; mC; mU; rC; rG; mC; rA; rU; mC; rG; rA; rG; LdT;rA$ mU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1436 3 10 17 5 zidB; rA; mC;rA; rA; rG; rA; mU; rA; mC; rU; mC; rG; mU; mC; mU; rG; rC; rG; rA; rG;rA; mC; rU; mC; rG; mC; mA; rU; mC; rG; rA; rG; LdT; rA$ rU; mU; rG; rU;zc3p; zc3p$ SERPINH1_6_S1437 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC;rU; mC; rG; LdT; rC; mU; rG; rC; rG; rA; rG; rA; mC; mU; rC; rG; mC; mA;rU; mC; rG; rA; rG; LdT; rA$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1438 3 15 17 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU;mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; rG; mC; mA;rU; mC; rG; rA; rG; LdT; rA$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1439 24 12 23 11 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC;rU; mC; rG; mU; rC; mU; rG; rC; rG; rA; rG; rA; mC; mU; rC; mG; rC; mA;rU; mC; rG; rA; rG; mU; rA; zc3p$ rU; mU; rG; mU; zc3p; zc3p$SERPINH1_6_S1442 24 29 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; mU; mC;rG; mU; mU; rG; rC; rG; rA; rG; rA; mC; mC; mU; rC; rG; mC; rA; rU; mC;rG; rA; rG; mU; rA; zc3p$ mU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S144324 9 22 7 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG; mU; mC;mU; rG; rC; rG; rA; rG; rA; mC; rU; mC; rG; mC; mA; rU; mC; rG; rA; rG;mU; rA; zc3p$ rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1444 zidB; rA; mC;rA; rA; rG; rA; mU; rA; mC; rU; mC; rG; LdT; rC; mU; rG; rC; rG; rA; rG;rA; mC; mU; rC; rG; mC; mA; rU; mC; rG; rA; rG; mU; rA; zc3p$ rU; mU;rG; rU; zc3p; zc3p$ SERPINH1_6_S1445 24 19 18 zidB; rA; mC; rA; rA; rG;rA; mU; rA; mC; rU; mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC;mU; rC; rG; mC; mA; rU; mC; rG; rA; rG; mU; rA; zc3p$ rU; mU; rG; rU;zc3p; zc3p$ SERPINH1_6_S1739 24 11 zidB; rA; mC; rA; rA; rG; rA; rU; mU;rA; mC; rU; mC; rG; rU2p; rG; rC; rG; rA; rG; rA; mC; rG; rC; mU; rC;mG; rC; mA; rU; mC; rA; rG; mU; rA; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1741 24 12 zidB; rA; mC; rA; rA; rG; rA; rU; mU; rA; mC; rU;mC; rG; rU2p; rG; rC; rG; rA; rG; rA; mC; rG; rC; mU; rC; rG; mC; mA;rU; rC; rA; rG; mU; rA; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1744 0 zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC;rG; rU2p; rG; rC; rG; rA; rG; rA; mC; rG; rC; mU; rC; rG; mC; mA; rU;mC; rA; rG; mU; rA; zc3p$ rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1746 0zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC; rG; rU2p; rG; rC;rG; rA; rG; rA; mC; rG; rC; mU; rC; mG; rC; mA; rU; mC; rA; rG; mU; rA;zc3p$ rU; mU; rG; mU; zc3p; zc3p$ SERPINH1_6_S1785 zidB; rA; mC; rA; rA;rG; rA; rU; dU; rA; mC; rU; mC; rG; rU2p; rC; rG; rC; rG; rA; rG; rA;mC; rG; mU; rC; rG; mC; mA; rU; rC; rU; rA; rG; mU; rA; zc3p$ mU; rG;rU; zc3p; zc3p$ SERPINH1_11_S1356 zidB; rC; rG; rG; rA; rC; rA; rG; mU;rG; mU; rU; mG; rU; mA; rG; rG; rC; rC; rU; rC; rU; rA; rC2p; mA; rG;mG; rC; mC; rU; mG; rA2p; rA2p; rC2p; rA2p rU; mC; rC; mG; zc3p; zc3p$SERPINH1_11_S1363 zc3p; rC; rG; rG; rA; rC; rA; rG; mU; rG; mU; rU; mG;rU; mA; rG; rG; rC; rC; rU; rC; rU; rA; rC2p; mA; rG; mG; rC; mC; rU;mG; rA2p; rA2p; rC2p; rA2p rU; mC; rC; mG; zc3p; zc3p$ SERPINH1_11_S1370zidB; rC; rG; rG; rA; rC; rA; rG; mU; rG; mU; rU; mG; rU; mA; rG; rG;rC; rC; rU; rC; rU; rA; rC2p; mA; rG; mG; rC; mC; rU; mG; rA2p; rA2p;rC2p; rA2p; zc3p$ rU; mC; rC; mG; zc3p; zc3p$ SERPINH1_11_S1446 zc3p;rC; rG; rG; rA; rC; rA; rG; mU; rG; mU; mU; rG; mU; rA; rG; rG; rC; rC;rU; rC; rU; rA; rC2p; rA; rG; rG; mC; mC; mU; rG; rA2p; rA2p; rC2p; rA2pmU; mC; mC; rG; zc3p; zc3p$ SERPINH1_11_S1449 zc3p; rC; rG; rG; rA; rC;rA; rG; mU; rG; mU; mU; rG; rU2p; rA; rG; rC; rC; rU; rC; rU; rA; rC2p;rG; rA; rG; rG; mC; rC; mU; rG; rU; rA2p; rA2p; rC2p; rA2p mC; mC; rG;zc3p; zc3p$ SERPINH1_11_S1450 zc3p; rC; rG; rG; rA; rC; rA; rG; mU; rG;mU; mU; rG; LdT; rA; rG; rG; rC; rC; rU; rC; rU; rA; rC2p; rA; rG; rG;mC; rC; mU; rG; rU; rA2p; rA2p; rC2p; rA2p mC; mC; rG; zc3p; zc3p$SERPINH1_11_S1451 zidB; rC; rG; rG; rA; rC; rA; rG; mU; rG; mU; mU; rG;mU; rA; rG; rG; rC; rC; rU; rC; rU; rA; rC2p; rA; rG; rG; mC; mC; mU;rG; rA2p; rA2p; rC2p; rA2p mU; mC; mC; rG; zc3p; zc3p$ SERPINH1_11_S1454zidB; rC; rG; rG; rA; rC; rA; rG; mU; rG; mU; mU; rG; rU2p; rA; rG; rC;rC; rU; rC; rU; rA; rC2p; rG; rA; rG; rG; mC; rC; mU; rG; rU; rA2p;rA2p; rC2p; rA2p mC; mC; rG; zc3p; zc3p$ SERPINH1_11_S1455 zidB; rC; rG;rG; rA; rC; rA; rG; mU; rG; mU; mU; rG; LdT; rA; rG; rG; rC; rC; rU; rC;rU; rA; rC2p; rA; rG; rG; mC; rC; mU; rG; rU; rA2p; rA2p; rC2p; rA2p mC;mC; rG; zc3p; zc3p$ SERPINH1_11_S1456 zidB; rC; rG; rG; rA; rC; rA; rG;mU; rG; mU; mU; rG; mU; rA; rG; rG; rC; rC; rU; rC; rU; rA; rC2p; rA;rG; rG; mC; mC; mU; rG; rA2p; rA2p; rC2p; rA2p; zc3p$ mU; mC; mC; rG;zc3p; zc3p$ SERPINH1_11_S1457 zidB; rC; rG; rG; rA; rC; rA; rG; mU; rG;mU; mU; rG; mU; rA; rG; rG; rC; rC; rU; rC; rU; rA; rC2p; rA; rG; rG;rC; mC; mU; rG; rA2p; rA2p; rC2p; rA2p; zc3p$ mU; mC; mC; rG; zc3p;zc3p$ SERPINH1_11_S1459 zidB; rC; rG; rG; rA; rC; rA; rG; mU; rG; mU;mU; rG; rU2p; rA; rG; rC; rC; rU; rC; rU; rA; rC2p; rG; rA; rG; rG; mC;rC; mU; rG; rU; rA2p; rA2p; rC2p; rA2p; zc3p$ mC; mC; rG; zc3p; zc3p$SERPINH1_11_S1460 zidB; rC; rG; rG; rA; rC; rA; rG; mU; rG; mU; mU; rG;LdT; rA; rG; rG; rC; rC; rU; rC; rU; rA; rC2p; rA; rG; rG; mC; rC; mU;rG; rU; rA2p; rA2p; rC2p; rA2p; zc3p$ mC; mC; rG; zc3p; zc3p$SERPINH1_11_S1461 zidB; mC; rG; rG; rA; mC; rA; mU; rG; mU; rU; mG; rU;mA; rG; rG; rG; rC; mC; rU; rC; mU; rA; mA; rG; mG; rC; mC; rU; mG; mC;rA; rA; LdC; rA$ rU; mC; rC; mG; zc3p; zc3p$ SERPINH1_11_S1462 zidB; mC;rG; rG; rA; mC; rA; mU; rG; mU; mU; rG; mU; rA; rG; rG; rG; rC; mC; rU;rC; mU; rA; rA; rG; rG; mC; mC; mU; rG; mC; rA; rA; LdC; rA$ mU; mC; mC;rG; zc3p; zc3p$ SERPINH1_11_S1464 45 43 zidB; mC; rG; rG; rA; mC; rA;mU; rG; mU; mU; rG; mU; rA; rG; rG; rG; rC; mC; rU; rC; mU; rA; rA; rG;rG; mC; rC; mU; rG; rU; mC; rA; rA; LdC; rA$ mC; mC; rG; zc3p; zc3p$SERPINH1_11_S1467 zidB; mC; rG; rG; rA; mC; rA; mU; rG; mU; rU; mG; rU;mA; rG; rG; rG; rC; rC; rU; rC; mU; rA; mA; rG; mG; rC; mC; rU; mG; mC;rA; rA; mC; rA; zc3p$ rU; mC; rC; mG; zc3p; zc3p$ SERPINH1_11_S1468zidB; mC; rG; rG; rA; mC; rA; mU; rG; mU; mU; rG; mU; rA; rG; rG; rG;rC; rC; rU; rC; mU; rA; rA; rG; rG; mC; mC; mU; rG; mC; rA; rA; mC; rA;zc3p$ mU; mC; mC; rG; zc3p; zc3p$ SERPINH1_11_S1469 zidB; mC; rG; rG;rA; mC; rA; mU; rG; mU; mU; rG; mU; rA; rG; rG; rG; rC; rC; rU; rC; mU;rA; rA; rG; rG; rC; mC; mU; rG; mC; rA; rA; mC; rA; zc3p$ mU; mC; mC;rG; zc3p; zc3p$ SERPINH1_11_S1470 zidB; mC; rG; rG; rA; mC; rA; mU; rG;mU; mU; rG; mU; rA; rG; rG; rG; rC; rC; rU; rC; mU; rA; rA; rG; rG; mC;rC; mU; rG; rU; mC; rA; rA; mC; rA; zc3p$ mC; mC; rG; zc3p; zc3p$SERPINH1_11_S1471 zidB; mC; rG; rG; rA; mC; rA; mU; rG; mU; mU; rG;rU2p; rA; rG; rG; rC; rC; rU; rC; mU; rA; rG; rA; rG; rG; mC; rC; mU;rG; rU; mC; rA; rA; mC; rA; zc3p$ mC; mC; rG; zc3p; zc3p$SERPINH1_11_S1472 zidB; mC; rG; rG; rA; mC; rA; mU; rG; mU; mU; rG; LdT;rA; rG; rG; rG; rC; rC; rU; rC; mU; rA; rA; rG; rG; mC; rC; mU; rG; rU;mC; rA; rA; mC; rA; zc3p$ mC; mC; rG; zc3p; zc3p$ SERPINH1_12_S1391zidB; rA; rC; rA; rA; rG; rA; rU; rA; mA; rC; mU; rC; mG; rU; mC; rG;rC; rG; rA; rG; rA; rC; rG2p; rU; rC; mG; rC; mA; rU; mC; rU; rA2p;rG2p; rU2p; rU2p mU; rG; mU; zc3p; zc3p$ SERPINH1_12_S1780 zidB; rA; mC;rA; mA; rG; rA; rA; rA; mC; rU; mC; rG; rU2p; rC; rU; rG; rC; rG; rA;rG; rA; mC; mU; rC; rG; mC; mA; rU; rC; rU; rG; rA; rG; mU; rU; zc3p$mU; rG; rU; zc3p; zc3p$ SERPINH1_30_S1391 zidB; rC; rG; rG; rA; rC; rA;rG; rA; mG; rU; mU; rG; mU; rA; mG; rG; rC; rC; rU; rC; rU; rA; rC2p;rA; rG; mG; rC; mC; rU; mG; rA2p; rA2p; rC2p; rU2p rU; mC; rC; mG; zc3p;zc3p$ SERPINH1_45_S1354 174 40 rA; rC; rU; rC; rC; rA; rA; rG; rA; yrA;rG; rG; rA; rA; rG; rU; rU; rG; rU; rC; rA; rA; rC; rU; rU; rC; rA; rU;rC; rU; rU; rG; rG; rA; rC; yrU; zdT; zdT$ rG; rU; zdT; zdT$SERPINH1_45_S1500 16 96 54 zidB; rA; rC; rU; rC; rC; rA; rA; ymA; rG;rG; rA; rA; rG; mU; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; rG; rA; mU;mC; mU; mU; rG; rU2p; rC2p; rC2p; yrU2p rG; rA; rG; rU; zc3p; zc3p$SERPINH1_45_S1501 zidB; rA; rC; rU; rC; rC; rA; rA; ymA; rG; mG; rA; mA;rG; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; mU; rC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p mG; rA; mG; rU; zc3p; zc3p$SERPINH1_45_S1502 zidB; rA; rC; rU; rC; rC; rA; rA; ymA; rG; mG; rA; mA;rG; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; rU; mC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p mG; rA; mG; rU; zc3p; zc3p$SERPINH1_45_S1505 16 22 17 zidB; rA; rC; rU; rC; rC; rA; rA; yrA; mG;rG; mA; rA; mG; rU; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; mU;rC; mU; rU; mG; rU2p; rC2p; rC2p; yrU2p rG; mA; rG; mU; zc3p; zc3p$SERPINH1_45_S1506 zc3p; rA; rC; rU; rC; rC; rA; rA; ymA; rG; rG; rA; rA;rG; mU; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; rG; rA; mU; mC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p rG; rA; rG; rU; zc3p; zc3p$SERPINH1_45_S1507 zc3p; rA; rC; rU; rC; rC; rA; rA; ymA; rG; mG; rA; mA;rG; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; mU; rC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p mG; rA; mG; rU; zc3p; zc3p$SERPINH1_45_S1508 zc3p; rA; rC; rU; rC; rC; rA; rA; ymA; rG; mG; rA; mA;rG; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; rU; mC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p mG; rA; mG; rU; zc3p; zc3p$SERPINH1_45_S1509 16 zc3p; rA; rC; rU; rC; rC; rA; rA; ymA; rG; rG; rA;rA; rG; rU2p; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; mU; rC; mU;mU; rG; rU2p; rC2p; rC2p; yrU2p mG; rA; mG; rU; zc3p; zc3p$SERPINH1_45_S1510 zc3p; rA; rC; rU; rC; rC; rA; rA; ymA; rG; rG; rA; rA;rG; LdT; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; rG; rA; mU; rC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p mG; rA; mG; rU; zc3p; zc3p$SERPINH1_45_S1511 8 27 zc3p; rA; rC; rU; rC; rC; rA; rA; yrA; mG; rG;mA; rA; mG; rU; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; mU; rC;mU; rU; mG; rU2p; rC2p; rC2p; yrU2p rG; mA; rG; mU; zc3p; zc3p$SERPINH1_45_S1512 zidB; rA; rC; rU; rC; rC; rA; rA; ymA; rG; rG; rA; rA;rG; mU; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; rG; rA; mU; mC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p; zc3p$ rG; rA; rG; rU; zc3p; zc3p$SERPINH1_45_S1513 zidB; rA; rC; rU; rC; rC; rA; rA; ymA; rG; mG; rA; mA;rG; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; mU; rC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p; zc3p$ mG; rA; mG; rU; zc3p; zc3p$SERPINH1_45_S1514 zidB; rA; rC; rU; rC; rC; rA; rA; ymA; rG; mG; rA; mA;rG; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; rU; mC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p; zc3p$ mG; rA; mG; rU; zc3p; zc3p$SERPINH1_45_S1515 zidB; rA; rC; rU; rC; rC; rA; rA; ymA; rG; rG; rA; rA;rG; rU2p; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; mU; rC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p; zc3p$ mG; rA; mG; rU; zc3p; zc3p$SERPINH1_45_S1516 zidB; rA; rC; rU; rC; rC; rA; rA; ymA; rG; rG; rA; rA;rG; LdT; mU; rG; rA; rU; rC; rA; rA; rC; rU2p; rG; rA; mU; rC; mU; mU;rG; rU2p; rC2p; rC2p; yrU2p; zc3p$ mG; rA; mG; rU; zc3p; zc3p$SERPINH1_45_S1517 24 22 31 7 11 14 zidB; rA; rC; rU; rC; rC; rA; rA;yrA; mG; rG; mA; rA; mG; rU; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG;rA; mU; rC; mU; rU; mG; rU2p; rC2p; rC2p; yrU2p; zc3p$ rG; mA; rG; mU;zc3p; zc3p$ SERPINH1_45_S1518 8 90 47 zidB; rA; rC; rU; mC; mC; rA; ymA;rG; rG; rA; rA; rG; mU; mU; rA; rG; rA; rU; mC; rA; rA; mC; rG; rA; mU;mC; mU; mU; rG; mU; rU; rC; LdC; yrU$ rG; rA; rG; rU; zc3p; zc3p$SERPINH1_45_S1523 3 17 30 16 zidB; rA; rC; rU; mC; mC; rA; yrA; mG; rG;mA; rA; mG; rU; rA; rG; rA; rU; mC; rA; rA; mC; mU; rG; rA; mU; rC; mU;rU; mG; mU; rU; rC; LdC; yrU$ rG; mA; rG; mU; zc3p; zc3p$SERPINH1_45_S1524 zidB; rA; rC; rU; rC; mC; rA; rA; ymA; rG; rG; rA; rA;rG; mU; mU; rG; rA; rU; mC; rA; rA; rC; rG; rA; mU; mC; mU; mU; rG; mU;rU; mC; mC; yrU; zc3p$ rG; rA; rG; rU; zc3p; zc3p$ SERPINH1_45_S1525zidB; rA; rC; rU; rC; mC; rA; rA; ymA; rG; mG; rA; mA; rG; mU; rG; rA;rU; mC; rA; rA; rC; mU; rG; rA; mU; rC; mU; mU; rG; mU; rU; mC; mC; yrU;zc3p$ mG; rA; mG; rU; zc3p; zc3p$ SERPINH1_45_S1529 24 17 33 zidB; rA;rC; rU; rC; mC; rA; rA; yrA; mG; rG; mA; rA; mG; rU; rG; rA; rU; mC; rA;rA; rC; mU; rG; rA; mU; rC; mU; rU; mG; mU; rU; mC; mC; yrU; zc3p$ rG;mA; rG; mU; zc3p; zc3p$ SERPINH1_45_S1684 24 14 zidB; rA; rC; rU; rC;rC; rA; rA; yrA; mG; rG; mA; rA; mG; rU2p; rG; rA; rU; rC; rA; rA; rC;rU2p; mU; rG; rA; mU; rC; mU; rU; rU2p; rC2p; rC2p; yrU2p; zc3p$ mG; rG;mA; rG; mU; zc3p; zc3p$ SERPINH1_45_S1685 8 15 zidB; rA; rC; rU; mC; mC;rA; yrA; mG; rG; mA; rA; mG; rU2p; rA; rG; rA; rU; mC; rA; rA; mC; mU;rG; rA; mU; rC; mU; rU; mU; rU; rC; LdC; yrU$ mG; rG; mA; rG; mU; zc3p;zc3p$ SERPINH1_45_S1781 zidB; rA; rC; rU; rC; rC; rA; rA; rU; rG; rG;mA; rA; mG; rU2p; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; mU; rC;mU; rU; mG; rU2p; rC2p; rC2p; yrU2p; zc3p$ rG; mA; rG; mU; zc3p; zc3p$SERPINH1_45_S1786 zidB; rA; rC; rU; rC; rC; rA; rA; dU; rG; rG; mA; rA;mG; rU2p; rG; rA; rU; rC; rA; rA; rC; rU2p; mU; rG; rA; mU; rC; mU; rU;mG; rU2p; rC2p; rC2p; rA2p; zc3p$ rG; mA; rG; mU; zc3p; zc3p$SERPINH1_51_S1356 zidB; rU; rC; rC; rU; rG; rA; rG; mU; rC; mA; rC; mC;rC; mA; rU; rA; rC; rA; rC; rA; rU; rG; rG2 mG; rU; mG; rU; mC; rU; mCSERPINH1_51_S1486 zidB; rU; rC; rC; rU; rG; rA; rG; mU; rC; rA; mC; rC;rC2p; rA; mU; rA; rC; rA; rC; rA; rU; rG; rG2p; rG; rU; rG; mU; mC; rU;mC; rG2p; rU2p; rG2p; rA2p; zc3p$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1487 zidB; rU; rC; rC; rU; rG; rA; rG; mU; rC; rA; mC; rC;LdC; rA; mU; rA; rC; rA; rC; rA; rU; rG; rG2p; rG; rU; rG; mU; mC; rU;mC; rG2p; rU2p; rG2p; rA2p; zc3p$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1488 zidB; rU; mC; rC; mU; rG; rA; mU; rC; mA; rC; mC; rC;mA; rU; rG; rA; rC; rA; mC; rA; mU; rG; mG; rU; mG; rU; mC; rU; mC; rG;rG; rU; LdG; rA$ rA; mG; rG; mA; zc3p; zc3p$ SERPINH1_51_S1489 8 25zidB; rU; mC; rC; mU; rG; rA; mU; mC; rA; mC; mC; mC; rA; rG; rA; rC;rA; mC; rA; mU; rG; mU; rG; mU; rG; mU; mC; mU; rG; rG; rU; LdG; rA$ mC;rA; rG; rG; rA; zc3p; zc3p$ SERPINH1_51_S1490 zidB; rU; mC; rC; mU; rG;rA; mU; rC; rA; mC; mC; mC; rA; mU; rG; rA; rC; rA; mC; rA; mU; rG; rG;rU; rG; mU; mC; mU; mC; rG; rG; rU; LdG; rA$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1491 zidB; rU; mC; rC; mU; rG; rA; mU; rC; rA; mC; mC; mC;rA; mU; rG; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG; mU; mC; rU; mC; rG;rG; rU; LdG; rA$ rA; rG; rG; rA; zc3p; zc3p$ SERPINH1_51_S1492 zidB; rU;mC; rC; mU; rG; rA; mU; rC; rA; mC; rC; rC2p; rA; mU; rG; rA; rC; rA;mC; rA; mU; rG; rG; rU; rG; mU; mC; rU; mC; rG; rG; rU; LdG; rA$ rA; rG;rG; rA; zc3p; zc3p$ SERPINH1_51_S1493 zidB; rU; mC; rC; mU; rG; rA; mU;rC; rA; mC; rC; LdC; rA; mU; rG; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG;mU; mC; rU; mC; rG; rG; rU; LdG; rA$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1494 zidB; rU; mC; rC; mU; rG; rA; mU; rC; mA; rC; mC; rC;mA; rU; rG; rA; rC; rA; mC; rA; mU; rG; mG; rU; mG; rU; mC; rU; mC; rG;rG; mU; rG; rA; zc3p$ rA; mG; rG; mA; zc3p; zc3p$ SERPINH1_51_S1495zidB; rU; mC; rC; mU; rG; rA; mU; mC; rA; mC; mC; mC; rA; rG; rA; rC;rA; mC; rA; mU; rG; mU; rG; mU; rG; mU; mC; mU; rG; rG; mU; rG; rA;zc3p$ mC; rA; rG; rG; rA; zc3p; zc3p$ SERPINH1_51_S1496 zidB; rU; mC;rC; mU; rG; rA; mU; rC; rA; mC; mC; mC; rA; mU; rG; rA; rC; rA; mC; rA;mU; rG; rG; rU; rG; mU; mC; mU; mC; rG; rG; mU; rG; rA; zc3p$ rA; rG;rG; rA; zc3p; zc3p$ SERPINH1_51_S1497 zidB; rU; mC; rC; mU; rG; rA; mU;rC; rA; mC; mC; mC; rA; mU; rG; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG;mU; mC; rU; mC; rG; rG; mU; rG; rA; zc3p$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1498 24 22 10 7 zidB; rU; mC; rC; mU; rG; rA; mU; rC; rA;mC; rC; rC2p; rA; mU; rG; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG; mU;mC; rU; mC; rG; rG; mU; rG; rA; zc3p$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1499 24 25 31 18 28 zidB; rU; mC; rC; mU; rG; rA; mU; rC;rA; mC; rC; LdC; rA; mU; rG; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG; mU;mC; rU; mC; rG; rG; mU; rG; rA; zc3p$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1666 zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; rA; mC; rC;rC2p; rA; mU; rA; rC2p; rA; mC; rA; mU; rG; rG; rU; rG; mU; mC; rU; mC;rG; rG; mU; rG; rA; zc3p$ rA; rG; rG; rA; zc3p; zc3p$ SERPINH1_51_S1667zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; rA; mC; rC; LdC; rA; mU; rA;rC2p; rA; mC; rA; mU; rG; rG; rU; rG; mU; mC; rU; mC; rG; rG; mU; rG;rA; zc3p$ rA; rG; rG; rA; zc3p; zc3p$ SERPINH1_51_S1668 16 14 zidB; rU;rC; rC; mU; rG; rA; rG; mU; rC; rA; mC; rC; rC2p; rA; mU; rA; rC2p; rA;mC; rA; mU; rG; rG; rU; rG; rU; mC; rU; mC; rA; rG; rG; mU; rG; rA;zc3p$ rG; rG; rA; zc3p; zc3p$ SERPINH1_51_S1669 24 18 zidB; rU; rC; rC;mU; rG; rA; rG; mU; rC; rA; mC; rC; rC2p; rA; mU; rA; rC2p; rA; mC; rA;mU; rG; rG; rU; mG; rU; rC; rU; mC; rA; rG; rG; mU; rG; rA; zc3p$ rG;rG; rA; zc3p; zc3p$ SERPINH1_51_S1670 16 13 zidB; rU; rC; rC; mU; rG;rA; rG; mU; rC; mA; rC; rC; rC2p; rA; mU; rA; rC2p; rA; mC; rA; mU; rG;rG; rU; rG; mU; mC; rU; mC; rG; rG; mU; rG; rA; zc3p$ rA; rG; rG; rA;zc3p; zc3p$ SERPINH1_51_S1673 24 22 zidB; rU; rC; rC; mU; rG; rA; rG;mU; rC; rA; mC; rC; rC2p; rA; mU; rA; rC; rA; mC; rA; mU; rG; rG; rU;rG; rU; mC; rU; mC; rA; rG; rG; mU; rG; rA; zc3p$ rG; rG; rA; zc3p;zc3p$ SERPINH1_51_S1674 16 zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; rA;mC; rC; rC2p; rA; mU; rA; rC; rA; mC; rA; mU; rG; rG; rU; mG; rU; rC;rU; mC; rA; rG; rG; mU; rG; rA; zc3p$ rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1675 16 35 zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; mA;rC; rC; rC2p; rA; mU; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG; mU; mC;rU; mC; rG; rG; mU; rG; rA; zc3p$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1676 10 zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; rA; mC;rC; rC2p; rA; mU; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG; mU; mC; rU;mC; rG; rG; rU2p; rG; rA; zc3p$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1677 10 zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; rA; mC;rC; LdC; rA; mU; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG; mU; mC; rU; mC;rG; rG; rU2p; rG; rA; zc3p$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1678 10 zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; rA; mC;rC; rC2p; rA; mU; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG; rU; mC; rU;mC; rA; rG; rG; rU2p; rG; rA; zc3p$ rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1679 10 zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; rA; mC;rC; rC2p; rA; mU; rA; rC; rA; mC; rA; mU; rG; rG; rU; mG; rU; rC; rU;mC; rA; rG; rG; rU2p; rG; rA; zc3p$ rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1680 10 zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; mA; rC;rC; rC2p; rA; mU; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG; mU; mC; rU;mC; rG; rG; rU2p; rG; rA; zc3p$ rA; rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1682 8 16 zidB; rU; mC; rC; mU; rG; rA; mU; rC; rA; mC; rC;rC2p; rA; mU; rG; rA; rC; rA; mC; rA; mU; rG; rG; rU; mG; rU; rC; rU;mC; rA; rG; rG; mU; rG; rA; zc3p$ rG; rG; rA; zc3p; zc3p$SERPINH1_51_S1778 zidB; rU; rC; rC; mU; rG; rA; rG; yrA; rC; rA; mC; rC;rC2p; rA; mU; rA; rC2p; rA; mC; rA; mU; rG; rG; rU; mG; rU; rC; rU; mC;rA; rG; rG; mU; rG; yrU; zc3p$ rG; rG; rA; zc3p; zc3p$ SERPINH1_51_S1779zidB; rU; mC; rC; mU; rG; rA; yrA; rC; rA; mC; rC; rC2p; rA; mU; rG; rA;rC; rA; mC; rA; mU; rG; rG; rU; mG; rU; rC; rU; mC; rA; rG; rG; mU; rG;yrU; zc3p$ rG; rG; rA; zc3p; zc3p$ SERPINH1_51_S1783 zidB; rU; rC; rC;mU; rG; rA; rG; dU; rC; rA; mC; rC; rC2p; rA; mU; rA; rC2p; rA; mC; rA;mU; rG; rG; rU; mG; rU; rC; rU; mC; rA; rG; rG; mU; rG; rA; zc3p$ rG;rG; rA; zc3p; zc3p$ SERPINH1_51_S1784 zidB; rU; mC; rC; mU; rG; rA; dU;rC; rA; mC; rC; rC2p; rA; mU; rG; rA; rC; rA; mC; rA; mU; rG; rG; rU;mG; rU; rC; rU; mC; rA; rG; rG; mU; rG; rA; zc3p$ rG; rG; rA; zc3p;zc3p$ SERPINH1_52_S1356 zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU;rC; mG; rU; mC; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA;mU; rC; mU; rG2p; rA2p; rG2p; rA2p rU; mG; rU; mC; zc3p; zc3p$SERPINH1_52_S1363 zc3p; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; rC; mG;rU; mC; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; mU; rC;mU; rG2p; rA2p; rG2p; rA2p rU; mG; rU; mC; zc3p; zc3p$ SERPINH1_52_S1370zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; rC; mG; rU; mC; rU; rU;rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; mU; rC; mU; rG2p; rA2p;rG2p; rA2p; zc3p$ rU; mG; rU; mC; zc3p; zc3p$ SERPINH1_52_S1552 zc3p;rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; mC; rG; mU; mC; rU; rG; rC; rG;rA; rG; rA; rC2p; mU; mC; rG; mC; rA; mU; mC; rG2p; rA2p; rG2p; rA2p mU;mU; rG; mU; rC; zc3p; zc3p$ SERPINH1_52_S1553 zc3p; rG; rA; rC; rA; rA;rG; rA; mU; rC; mU; mC; rG; mU; mC; rU; rU; rG; rC; rG; rA; rG; rA;rC2p; mC; rG; mC; rA; mU; mC; rU; rG2p; rA2p; rG2p; rA2p; mU; rG; mU;rC; zc3p; zc3p$ SERPINH1_52_S1554 zc3p; rG; rA; rC; rA; rA; rG; rA; mU;rC; mU; mC; rG; mU; mC; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG;mC; rA; rU; mC; mU; rG2p; rA2p; rG2p; rA2p mU; rG; mU; rC; zc3p; zc3p$SERPINH1_52_S1555 zc3p; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; mC; rG;mU; mC; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; mU; rC;mU; rG2p; rA2p; rG2p; rA2p mU; rG; mU; rC; zc3p; zc3p$ SERPINH1_52_S1556zc3p; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; mC; rG; rU; LdC; rU; rU;rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; rU; mC; mU; rG2p; rA2p;rG2p; rA2p mU; rG; mU; rC; zc3p; zc3p$ SERPINH1_52_S1557 zc3p; rG; rA;rC; rA; rA; rG; rA; mU; rC; mU; mC; rG; rU; rC2p; rU; rU; rG; rC; rG;rA; rG; rA; rC2p; mC; rG; mC; rA; rU; mC; mU; rG2p; rA2p; rG2p; rA2p mU;rG; mU; rC; zc3p; zc3p$ SERPINH1_52_S1558 zidB; rG; rA; rC; rA; rA; rG;rA; mU; rC; mU; mC; rG; mU; mC; rU; rG; rC; rG; rA; rG; rA; rC2p; mU;mC; rG; mC; rA; mU; mC; rG2p; rA2p; rG2p; rA2p mU; mU; rG; mU; rC; zc3p;zc3p$ SERPINH1_52_S1559 zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU;mC; rG; mU; mC; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA;mU; mC; rU; rG2p; rA2p; rG2p; rA2p mU; rG; mU; rC; zc3p; zc3p$SERPINH1_52_S1560 zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; mC; rG;mU; mC; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; rU; mC;mU; rG2p; rA2p; rG2p; rA2p mU; rG; mU; rC; zc3p; zc3p$ SERPINH1_52_S1561zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; mC; rG; mU; mC; rU; rU;rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; mU; rC; mU; rG2p; rA2p;rG2p; rA2p mU; rG; mU; rC; zc3p; zc3p$ SERPINH1_52_S1562 zidB; rG; rA;rC; rA; rA; rG; rA; mU; rC; mU; mC; rG; rU; LdC; rU; rU; rG; rC; rG; rA;rG; rA; rC2p; mC; rG; mC; rA; rU; mC; mU; rG2p; rA2p; rG2p; rA2p mU; rG;mU; rC; zc3p; zc3p$ SERPINH1_52_S1563 zidB; rG; rA; rC; rA; rA; rG; rA;mU; rC; mU; mC; rG; rU; rC2p; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC;rG; mC; rA; rU; mC; mU; rG2p; rA2p; rG2p; rA2p mU; rG; mU; rC; zc3p;zc3p$ SERPINH1_52_S1564 16 94 zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC;mU; mC; rG; mU; mC; rU; rG; rC; rG; rA; rG; rA; rC2p; mU; mC; rG; mC;rA; mU; mC; rG2p; rA2p; rG2p; rA2p; zc3p$ mU; mU; rG; mU; rC; zc3p;zc3p$ SERPINH1_52_S1565 zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU;mC; rG; mU; mC; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA;mU; mC; rU; rG2p; rA2p; rG2p; rA2p; zc3p$ mU; rG; mU; rC; zc3p; zc3p$SERPINH1_52_S1566 zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; mC; rG;mU; mC; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; rU; mC;mU; rG2p; rA2p; rG2p; rA2p; zc3p$ mU; rG; mU; rC; zc3p; zc3p$SERPINH1_52_S1567 zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; mC; rG;mU; mC; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; mU; rC;mU; rG2p; rA2p; rG2p; rA2p; zc3p$ mU; rG; mU; rC; zc3p; zc3p$SERPINH1_52_S1568 zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; mC; rG;rU; LdC; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; rU; mC;mU; rG2p; rA2p; rG2p; rA2p; zc3p$ mU; rG; mU; rC; zc3p; zc3p$SERPINH1_52_S1569 zidB; rG; rA; rC; rA; rA; rG; rA; mU; rC; mU; mC; rG;rU; rC2p; rU; rU; rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; rU; mC;mU; rG2p; rA2p; rG2p; rA2p; zc3p$ mU; rG; mU; rC; zc3p; zc3p$SERPINH1_52_S1570 zidB; rG; rA; mC; rA; rA; rG; rA; mU; rC; mU; rC; mG;rU; mC; rU; mU; rG; mC; rG; rA; rG; rA; mC; rG; mC; rA; mU; rC; mU; mC;rG; rA; LdG; rA$ rU; mG; rU; mC; zc3p; zc3p$ SERPINH1_52_S1571 zidB; rG;rA; mC; rA; rA; rG; rA; mU; rC; mU; mC; rG; mU; mC; mU; rG; mC; rG; rA;rG; rA; mU; mC; rG; mC; rA; mU; mC; mC; rG; rA; LdG; rA$ mU; mU; rG; mU;rC; zc3p; zc3p$ SERPINH1_52_S1572 zidB; rG; rA; mC; rA; rA; rG; rA; mU;rC; mU; mC; rG; mU; mC; rU; mU; rG; mC; rG; rA; rG; rA; mC; rG; mC; rA;mU; mC; rU; mC; rG; rA; LdG; rA$ mU; rG; mU; rC; zc3p; zc3p$SERPINH1_52_S1573 zidB; rG; rA; mC; rA; rA; rG; rA; mU; rC; mU; mC; rG;mU; mC; rU; mU; rG; mC; rG; rA; rG; rA; mC; rG; mC; rA; rU; mC; mU; mC;rG; rA; LdG; rA$ mU; rG; mU; rC; zc3p; zc3p$ SERPINH1_52_S1574 zidB; rG;rA; mC; rA; rA; rG; rA; mU; rC; mU; mC; rG; mU; mC; rU; mU; rG; mC; rG;rA; rG; rA; mC; rG; mC; rA; mU; rC; mU; mC; rG; rA; LdG; rA$ mU; rG; mU;rC; zc3p; zc3p$ SERPINH1_52_S1575 zidB; rG; rA; mC; rA; rA; rG; rA; mU;rC; mU; mC; rG; rU; LdC; rU; mU; rG; mC; rG; rA; rG; rA; mC; rG; mC; rA;rU; mC; mU; mC; rG; rA; LdG; rA$ mU; rG; mU; rC; zc3p; zc3p$SERPINH1_52_S1576 zidB; rG; rA; mC; rA; rA; rG; rA; mU; rC; mU; mC; rG;rU; rC2p; rU; mU; rG; mC; rG; rA; rG; rA; mC; rG; mC; rA; rU; mC; mU;mC; rG; rA; LdG; rA$ mU; rG; mU; rC; zc3p; zc3p$ SERPINH1_52_S1577 zidB;rG; rA; mC; rA; rA; rG; rA; mU; rC; mU; rC; mG; rU; mC; rU; mU; rG; mC;rG; rA; rG; rA; mC; rG; mC; rA; mU; rC; mU; mC; rG; rA; rG; rA; zc3p$rU; mG; rU; mC; zc3p; zc3p$ SERPINH1_52_S1578 zidB; rG; rA; mC; rA; rA;rG; rA; mU; rC; mU; mC; rG; mU; mC; mU; rG; mC; rG; rA; rG; rA; mU; mC;rG; mC; rA; mU; mC; mC; rG; rA; rG; rA; zc3p$ mU; mU; rG; mU; rC; zc3p;zc3p$ SERPINH1_52_S1579 zidB; rG; rA; mC; rA; rA; rG; rA; mU; rC; mU;mC; rG; mU; mC; rU; mU; rG; mC; rG; rA; rG; rA; mC; rG; mC; rA; mU; mC;rU; mC; rG; rA; rG; rA; zc3p$ mU; rG; mU; rC; zc3p; zc3p$SERPINH1_52_S1580 zidB; rG; rA; mC; rA; rA; rG; rA; mU; rC; mU; mC; rG;mU; mC; rU; mU; rG; mC; rG; rA; rG; rA; mC; rG; mC; rA; rU; mC; mU; mC;rG; rA; rG; rA; zc3p$ mU; rG; mU; rC; zc3p; zc3p$ SERPINH1_52_S1581zidB; rG; rA; mC; rA; rA; rG; rA; mU; rC; mU; mC; rG; mU; mC; rU; mU;rG; mC; rG; rA; rG; rA; mC; rG; mC; rA; mU; rC; mU; mC; rG; rA; rG; rA;zc3p$ mU; rG; mU; rC; zc3p; zc3p$ SERPINH1_52_S1582 zidB; rG; rA; mC;rA; rA; rG; rA; mU; rC; mU; mC; rG; rU; LdC; rU; mU; rG; mC; rG; rA; rG;rA; mC; rG; mC; rA; rU; mC; mU; mC; rG; rA; rG; rA; zc3p$ mU; rG; mU;rC; zc3p; zc3p$ SERPINH1_52_S1583 zidB; rG; rA; mC; rA; rA; rG; rA; mU;rC; mU; mC; rG; rU; rC2p; rU; mU; rG; mC; rG; rA; rG; rA; mC; rG; mC;rA; rU; mC; mU; mC; rG; rA; rG; rA; zc3p$ mU; rG; mU; rC; zc3p; zc3p$SERPINH1_58_S1391 zidB; rG; rA; rC; rA; rA; rG; rA; rA; mC; rU; mC; rG;mU; rC; mU; rU; rG; rC; rG; rA; rG; rA; rC2p; rC; rG; mC; rA; mU; rC;mU; rU; rG2p; rA2p; rG2p; rU2p mG; rU; mC; zc3p; zc3p$ SERPINH1_58_S1584zidB; rG; rA; rC; rA; rA; rG; rA; rA; mC; rU; mC; rG; mU; mC; rU; rU;rG; rC; rG; rA; rG; rA; rC2p; mC; rG; mC; rA; rU; mC; mU; rG2p; rA2p;rG2p; rU2p mU; rG; mU; rC; zc3p; zc3p$ SERPINH1_86_S1356 16 68 65 zidB;rA; rC; rA; rG; rG; rC; rC; mU; rG; mU; rA; mG; rU; mU; rG; rU; rC; rU;rA; rC; rA; rA; rC2p; mU; rA; mG; rA; mG; rG; mC; rU2p; rA2p; rC2p; rA2prC; mU; rG; mU; zc3p; zc3p$ SERPINH1_86_S1363 zc3p; rA; rC; rA; rG; rG;rC; rC; mU; rG; mU; rA; mG; rU; mU; rG; rU; rC; rU; rA; rC; rA; rA;rC2p; mU; rA; mG; rA; mG; rG; mC; rU2p; rA2p; rC2p; rA2p rC; mU; rG; mU;zc3p; zc3p$ SERPINH1_86_S1370 zidB; rA; rC; rA; rG; rG; rC; rC; mU; rG;mU; rA; mG; rU; mU; rG; rU; rC; rU; rA; rC; rA; rA; rC2p; mU; rA; mG;rA; mG; rG; mC; rU2p; rA2p; rC2p; rA2p; zc3p$ rC; mU; rG; mU; zc3p;zc3p$ SERPINH1_86_S1530 zc3p; rA; rC; rA; rG; rG; rC; rC; mU; rG; mU;rA; rG; mU; mU; rG; rU; rC; rU; rA; rC; rA; rA; rC2p; mU; rA; rG; rA;rG; rG; mC; rU2p; rA2p; rC2p; rA2p mC; mU; rG; rU; zc3p; zc3p$SERPINH1_86_S1531 52 31 zc3p; rA; rC; rA; rG; rG; rC; rC; mU; rG; mU;rA; rG; mU; mU; rG; rU; rC; rU; rA; rC; rA; rA; rC2p; mU; rA; rG; rA;rG; rG; mC; rC; rU2p; rA2p; rC2p; rA2p mU; rG; rU; zc3p; zc3p$SERPINH1_86_S1532 zc3p; rA; rC; rA; rG; rG; rC; rC; mU; rG; mU; rA; rG;rU; LdT; rG; rU; rC; rU; rA; rC; rA; rA; rC2p; mU; rA; rG; rA; rG; rG;mC; rU2p; rA2p; rC2p; rA2p mC; mU; rG; rU; zc3p; zc3p$ SERPINH1_86_S15338 70 74 zc3p; rA; rC; rA; rG; rG; rC; rC; mU; rG; mU; rA; rG; rU; rU2p;rG; rU; rC; rU; rA; rC; rA; rA; rC2p; mU; rA; rG; rA; rG; rG; mC; rU2p;rA2p; rC2p; rA2p mC; mU; rG; rU; zc3p; zc3p$ SERPINH1_86_S1534 zidB; rA;rC; rA; rG; rG; rC; rC; mU; rG; mU; rA; rG; mU; mU; rG; rU; rC; rU; rA;rC; rA; rA; rC2p; mU; rA; rG; rA; rG; rG; mC; rU2p; rA2p; rC2p; rA2p mC;mU; rG; rU; zc3p; zc3p$ SERPINH1_86_S1535 zidB; rA; rC; rA; rG; rG; rC;rC; mU; rG; mU; rA; rG; mU; mU; rG; rU; rC; rU; rA; rC; rA; rA; rC2p;mU; rA; rG; rA; rG; rG; mC; rC; rU2p; rA2p; rC2p; rA2p mU; rG; rU; zc3p;zc3p$ SERPINH1_86_S1536 zidB; rA; rC; rA; rG; rG; rC; rC; mU; rG; mU;rA; rG; rU; LdT; rG; rU; rC; rU; rA; rC; rA; rA; rC2p; mU; rA; rG; rA;rG; rG; mC; rU2p; rA2p; rC2p; rA2p mC; mU; rG; rU; zc3p; zc3p$SERPINH1_86_S1537 zidB; rA; rC; rA; rG; rG; rC; rC; mU; rG; mU; rA; rG;rU; rU2p; rG; rU; rC; rU; rA; rC; rA; rA; rC2p; mU; rA; rG; rA; rG; rG;mC; rU2p; rA2p; rC2p; rA2p mC; mU; rG; rU; zc3p; zc3p$ SERPINH1_86_S1538zidB; rA; rC; rA; rG; rG; rC; rC; mU; rG; mU; rA; rG; mU; mU; rG; rU;rC; rU; rA; rC; rA; rA; rC2p; mU; rA; rG; rA; rG; rG; mC; rU2p; rA2p;rC2p; rA2p; zc3p$ mC; mU; rG; rU; zc3p; zc3p$ SERPINH1_86_S1539 zidB;rA; rC; rA; rG; rG; rC; rC; mU; rG; mU; rA; rG; mU; mU; rG; rU; rC; rU;rA; rC; rA; rA; rC2p; mU; rA; rG; rA; rG; rG; mC; rC; rU2p; rA2p; rC2p;rA2p; zc3p$ mU; rG; rU; zc3p; zc3p$ SERPINH1_86_S1540 zidB; rA; rC; rA;rG; rG; rC; rC; mU; rG; mU; rA; rG; rU; LdT; rG; rU; rC; rU; rA; rC; rA;rA; rC2p; mU; rA; rG; rA; rG; rG; mC; rU2p; rA2p; rC2p; rA2p; zc3p$ mC;mU; rG; rU; zc3p; zc3p$ SERPINH1_86_S1541 zidB; rA; rC; rA; rG; rG; rC;rC; mU; rG; mU; rA; rG; rU; rU2p; rG; rU; rC; rU; rA; rC; rA; rA; rC2p;mU; rA; rG; rA; rG; rG; mC; rU2p; rA2p; rC2p; rA2p; zc3p$ mC; mU; rG;rU; zc3p; zc3p$ SERPINH1_86_S1542 zidB; rA; mC; rA; rG; rG; mC; mU; rG;mU; rA; mG; rU; mU; rG; rC; rU; rC; mU; rA; mC; rA; rA; mU; rA; mG; rA;mG; rG; mC; rC; mU; rA; LdC; rA$ rC; mU; rG; mU; zc3p; zc3p$SERPINH1_86_S1543 8 44 42 zidB; rA; mC; rA; rG; rG; mC; mU; rG; mU; rA;rG; mU; mU; rG; rC; rU; rC; mU; rA; mC; rA; rA; mU; rA; rG; rA; rG; rG;mC; rC; mU; rA; LdC; rA$ mC; mU; rG; rU; zc3p; zc3p$ SERPINH1_86_S1544 829 36 zidB; rA; mC; rA; rG; rG; mC; mU; rG; mU; rA; rG; mU; mU; rG; rC;rU; rC; mU; rA; mC; rA; rA; mU; rA; rG; rA; rG; rG; mC; rC; rC; mU; rA;LdC; rA$ mU; rG; rU; zc3p; zc3p$ SERPINH1_86_S1545 zidB; rA; mC; rA; rG;rG; mC; mU; rG; mU; rA; rG; rU; LdT; rG; rC; rU; rC; mU; rA; mC; rA; rA;mU; rA; rG; rA; rG; rG; mC; rC; mU; rA; LdC; rA$ mC; mU; rG; rU; zc3p;zc3p$ SERPINH1_86_S1546 16 67 63 zidB; rA; mC; rA; rG; rG; mC; mU; rG;mU; rA; rG; rU; rU2p; rG; rC; rU; rC; mU; rA; mC; rA; rA; mU; rA; rG;rA; rG; rG; mC; rC; mU; rA; LdC; rA$ mC; mU; rG; rU; zc3p; zc3p$SERPINH1_86_S1547 16 24 63 zidB; rA; mC; rA; rG; rG; rC; rC; mU; rG; mU;rA; mG; rU; mU; rG; rU; rC; mU; rA; mC; rA; rA; mU; rA; mG; rA; mG; rG;mC; rC; mU; rA; mC; rA; zc3p$ rC; mU; rG; mU; zc3p; zc3p$SERPINH1_86_S1548 16 39 67 zidB; rA; mC; rA; rG; rG; rC; rC; mU; rG; mU;rA; rG; mU; mU; rG; rU; rC; mU; rA; mC; rA; rA; mU; rA; rG; rA; rG; rG;mC; rC; mU; rA; mC; rA; zc3p$ mC; mU; rG; rU; zc3p; zc3p$SERPINH1_86_S1549 16 20 68 zidB; rA; mC; rA; rG; rG; rC; rC; mU; rG; mU;rA; rG; mU; mU; rG; rU; rC; mU; rA; mC; rA; rA; mU; rA; rG; rA; rG; rG;mC; rC; rC; mU; rA; mC; rA; zc3p$ mU; rG; rU; zc3p; zc3p$SERPINH1_86_S1550 16 96 92 zidB; rA; mC; rA; rG; rG; rC; rC; mU; rG; mU;rA; rG; rU; LdT; rG; rU; rC; mU; rA; mC; rA; rA; mU; rA; rG; rA; rG; rG;mC; rC; mU; rA; mC; rA; zc3p$ mC; mU; rG; rU; zc3p; zc3p$SERPINH1_86_S1551 16 70 51 zidB; rA; mC; rA; rG; rG; rC; rC; mU; rG; mU;rA; rG; rU; rU2p; rG; rU; rC; mU; rA; mC; rA; rA; mU; rA; rG; rA; rG;rG; mC; rC; mU; rA; mC; rA; zc3p$ mC; mU; rG; rU; zc3p; zc3p$SERPINH1_2_S1686 zidB; rG; rA; rG; rA; rC; rA; mC; mU; rA; mU; rA; mG;rC; mA; rC; rA; rU; rG; rG; rG; mU; rG; rC; mC; rC; mA; rU; mG; rU; mG;mU; rA; mU; rA; zc3p$ rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1688 zidB;rG; rA; rG; rA; rC; rA; mC; mU; rA; mU; rA; mG; rC; rA2p; rA; rU; rG;rG; rG; mU; rG; rC; rC; mC; rC; mA; rU; mG; rU; mG; mU; rA; mU; rA;zc3p$ rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1689 zidB; rG; rA; rG; rA;rC; rA; mC; mU; rA; mU; rA; mG; rC; mA; rC; rA; rU; rG; rG; rG; mU; rG;rC; mC; rC; mA; rU; mG; rU; mG; mU; rA; mU; rA; zc3p; zc3p$ rU; mC; rU;mC; zc3p; zc3p$ SERPINH1_2_S1690 zidB; rG; rA; rG; rA; rC; rA; mC; mU;rA; mU; rA; mG; rC; mA; rC; rA; rU; rG; rG; rG; mU; rG; rC; mC; rC; mA;rU; mG; rU; mG; mU; rA; mU; rA; zc3p; zc3p$ rU; mC; rU; rC; zc3p; zc3p$SERPINH1_2_S1691 zidB; rG; rA; rG; rA; rC; rA; mC; mU; rA; mU; rA; mG;rC; rA2p; rA; rU; rG; rG; rG; mU; rG; rC; rC; mC; rC; mA; rU; mG; rU;mG; mU; rA; mU; rA; zc3p; zc3p$ rU; mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1692 zidB; rG; rA; rG; rA; rC; rA; mC; mU; rA; mU; rA; mG;rC; mA; rC; rA; rU2p; rG; rG; rG; mU; rG; mC; rC; mA; rU; mG; rU; mG;rC; mU; rA; mU; rA; zc3p$ rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1693zidB; rG; rA; rG; rA; rC; rA; mC; mU; rA; mU; rA; mG; rC; mA; rC; rA;rU2p; rG; rG; rG; mU; rG; mC; rC; mA; rU; mG; rU; mG; rC; mU; rA; mU;rA; zc3p$ rU; mC; rU; rC; zc3p; zc3p$ SERPINH1_2_S1695 zidB; rG; rA; rG;rA; mC; rA; mU; rA; mU; rA; mG; rC; mA; rC; mC; rA; rU; rG; rG; rG; rU;rG; mC; rC; mA; rU; mG; rU; mG; rC; mU; rA; mU; rA; zc3p$ rU; mC; rU;mC; zc3p; zc3p$ SERPINH1_2_S1696 zidB; rG; rA; rG; rA; mC; rA; mU; rA;mU; rA; mG; rC; mA; rC; mC; rA; rU; rG; rG; rG; rU; rG; mC; rC; mA; rU;mG; rU; mG; rC; mU; rA; mU; rA; zc3p$ rU; mC; rU; rC; zc3p; zc3p$SERPINH1_2_S1697 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; mG; rC;rA2p; mC; rA; rU; rG; rG; rG; rU; rG; rC; mC; rC; mA; rU; mG; rU; mG;rC; mU; rA; mU; rA; zc3p$ rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1698zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; mG; rC; mA; rC; mC; rA;rU; rG; rG; rG; mU; rG; mC; rC; mA; rU; mG; rU; mG; rC; mU; rA; rU2p;rA; zc3p$ rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1699 zidB; rG; rA; rG;rA; mC; rA; mU; rA; mU; rA; mG; rC; mA; rC; mC; rA; rU; rG; rG; rG; mU;rG; mC; rC; mA; rU; mG; rU; mG; rC; mU; rA; rU2p; rA; zc3p$ rU; mC; rU;rC; zc3p; zc3p$ SERPINH1_2_S1701 zidB; rG; rA; rG; rA; mC; rA; mU; rA;mU; rA; mG; rC; mA; rC; mC; rA; rU; rG; rG; rG; mU; rG; mC; rC; mA; rU;mG; rU; mG; rC; mU; rA; LdT; rA; zc3p$ rU; mC; rU; mC; zc3p; zc3p$SERPINH1_2_S1702 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; mG; rC;mA; rC; mC; rA; rU; rG; rG; rG; mU; rG; mC; rC; mA; rU; mG; rU; mG; rC;mU; rA; LdT; rA; zc3p$ rU; mC; rU; rC; zc3p; zc3p$ SERPINH1_2_S1703zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; mG; rC; rA2p; mC; rA; rU;rG; rG; rG; mU; rG; rC; mC; rC; mA; rU; mG; rU; mG; rC; mU; rA; LdT; rA;zc3p$ rU; mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1704 zidB; rG; rA; rG; rA;rC; rA; rC; mU; rA; mU; rA; mG; rC; mA; rC; rA; rU; rG; rG; rG; rU; rG;rC2p; mC; rC; mA; rU; mG; rU; mG; rU2p; rA2p; rU2p; rA2p rU; mC; rU; rC;zc3p; zc3p$ SERPINH1_2_S1706 zidB; rG; rA; rG; rA; rC; rA; rC; mU; rA;mU; rA; mG; rC; mA; rC; rA; rU; rG; rG; rG; rU; rG; rC2p; mC; rC; mA;rU; mG; rU; mG; rU2p; rA2p; rU2p; rA2p; zc3p$ rU; mC; rU; rC; zc3p;zc3p$ SERPINH1_2_S1708 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; mG;rC; mA; rC; mC; rA; rU; rG; rG; rG; mU; rG; mC; rC; mA; rU; mG; rU; mG;mC; mU; rA; LdT; rA$ rU; mC; rU; rC; zc3p; zc3p$ SERPINH1_2_S1709 zidB;rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; mG; rC; rA2p; mC; rA; rU; rG;rG; rG; mU; rG; rC; mC; rC; mA; rU; mG; rU; mG; mC; mU; rA; LdT; rA$ rU;mC; rU; mC; zc3p; zc3p$ SERPINH1_2_S1710 zidB; rG; rA; rG; rA; mC; rA;mU; rA; mU; rA; mG; rC; mA; rC; mC; rA; rU; rG; rG; rG; mU; rG; mC; rC;mA; rU; mG; rU; mG; rC; mU; rA; mU; rA; zc3p$ rU; mC; rU; rC; zc3p;zc3p$ SERPINH1_2_S1711 zidB; rG; rA; rG; rA; mC; rA; mU; rA; mU; rA; mG;rC; rA2p; mC; rA; rU; rG; rG; rG; mU; rG; rC; mC; rC; mA; rU; mG; rU;mG; rC; mU; rA; mU; rA; zc3p$ rU; mC; rU; mC; zc3p; zc3p$SERPINH1_6_S1712 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG;mU; rC; mU; rG; rC; rG; rA; rG; rA; mC; mU; rC; mG; rC; mA; rU; mC; rG;rA; rG; LdT; rA; zc3p$ rU; mU; rG; mU; zc3p; zc3p$ SERPINH1_6_S1713zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG; mU; mC; mU; rG;rC; rG; rA; rG; rA; mC; rU; mC; rG; mC; mA; rU; mC; rG; rA; rG; LdT; rA;zc3p$ rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1714 zidB; rA; mC; rA; rA;rG; rA; mU; rA; mC; rU; mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC;rC; mU; rC; rG; mC; mA; rU; mC; rG; rA; rG; LdT; rA; zc3p$ rU; mU; rG;rU; zc3p; zc3p$ SERPINH1_6_S1715 zidB; rA; mC; rA; rA; rG; rA; mU; rA;mC; rU; mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; mG;rC; mA; rU; mC; rG; rA; rG; LdT; rA; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1716 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG;rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; mG; rC; mA; rU; mC;rG; rA; rG; LdT; rA; zc3p$ rU; mU; rG; mU; zc3p; zc3p$ SERPINH1_6_S1717zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG; rU2p; mU; rG; rC;rG; rA; rG; rA; mC; rC; mU; rC; rG; mC; mA; rU; rC; rG; rA; rG; LdT; rA;zc3p$ rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1718 zidB; rA; mC; rA; rA;rG; rA; mU; rA; mC; rU; mC; rG; mU; rC; mU; rG; rC; rG; rA; rG; rA; mC;mU; rC; mG; rC; mA; rU; mC; rG; rA; rG; LdT; rA; zc3p; zc3p$ rU; mU; rG;mU; zc3p; zc3p$ SERPINH1_6_S1719 zidB; rA; mC; rA; rA; rG; rA; mU; rA;mC; rU; mC; rG; mU; mC; mU; rG; rC; rG; rA; rG; rA; mC; rU; mC; rG; mC;mA; rU; mC; rG; rA; rG; LdT; rA; zc3p; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1720 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG;rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; rG; mC; mA; rU; mC;rG; rA; rG; LdT; rA; zc3p; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1721 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG;rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; mG; rC; mA; rU; mC;rG; rA; rG; LdT; rA; zc3p; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1722 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG;rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; mG; rC; mA; rU; mC;rG; rA; rG; LdT; rA; zc3p; zc3p$ rU; mU; rG; mU; zc3p; zc3p$SERPINH1_6_S1723 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG;rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; rG; mC; mA; rU; rC;rG; rA; rG; LdT; rA; zc3p; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1724 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG;mU; rC; mU; rG; rC; rG; rA; rG; rA; mC; mU; rC; mG; rC; mA; rU; mC; rG;rA; rG; rU2p; rA; zc3p$ rU; mU; rG; mU; zc3p; zc3p$ SERPINH1_6_S1725zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG; mU; mC; mU; rG;rC; rG; rA; rG; rA; mC; rU; mC; rG; mC; mA; rU; mC; rG; rA; rG; rU2p;rA; zc3p$ rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1726 zidB; rA; mC; rA;rA; rG; rA; mU; rA; mC; rU; mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA;mC; rC; mU; rC; rG; mC; mA; rU; mC; rG; rA; rG; rU2p; rA; zc3p$ rU; mU;rG; rU; zc3p; zc3p$ SERPINH1_6_S1727 zidB; rA; mC; rA; rA; rG; rA; mU;rA; mC; rU; mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC;mG; rC; mA; rU; mC; rG; rA; rG; rU2p; rA; zc3p$ rU; mU; rG; rU; zc3p;zc3p$ SERPINH1_6_S1728 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC;rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; mG; rC; mA; rU;mC; rG; rA; rG; rU2p; rA; zc3p$ rU; mU; rG; mU; zc3p; zc3p$SERPINH1_6_S1729 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG;rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; rG; mC; mA; rU; rC;rG; rA; rG; rU2p; rA; zc3p$ rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1730zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG; mU; rC; mU; rG;rC; rG; rA; rG; rA; mC; mU; rC; mG; rC; mA; rU; mC; rG; rA; rG; rU2p;rA; zc3p; zc3p$ rU; mU; rG; mU; zc3p; zc3p$ SERPINH1_6_S1731 zidB; rA;mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG; mU; mC; mU; rG; rC; rG; rA;rG; rA; mC; rU; mC; rG; mC; mA; rU; mC; rG; rA; rG; rU2p; rA; zc3p;zc3p$ rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1732 zidB; rA; mC; rA; rA;rG; rA; mU; rA; mC; rU; mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC;rC; mU; rC; rG; mC; mA; rU; mC; rG; rA; rG; rU2p; rA; zc3p; zc3p$ rU;mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1733 zidB; rA; mC; rA; rA; rG; rA;mU; rA; mC; rU; mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU;rC; mG; rC; mA; rU; mC; rG; rA; rG; rU2p; rA; zc3p; zc3p$ rU; mU; rG;rU; zc3p; zc3p$ SERPINH1_6_S1734 zidB; rA; mC; rA; rA; rG; rA; mU; rA;mC; rU; mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; mG;rC; mA; rU; mC; rG; rA; rG; rU2p; rA; zc3p; zc3p$ rU; mU; rG; mU; zc3p;zc3p$ SERPINH1_6_S1735 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC;rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; rG; mC; mA; rU;rC; rG; rA; rG; rU2p; rA; zc3p; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1736 zidB; rA; mC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC;rG; mU; rC; rG; rC; rG; rA; rG; rA; mC; rG; mU; rC; mG; rC; mA; rU; mC;rA; rG; mU; rA; zc3p$ rU; mU; rG; mU; zc3p; zc3p$ SERPINH1_6_S1737 zidB;rA; mC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC; rG; mU; mC; rG; rC; rG;rA; rG; rA; mC; rG; rU; mC; rG; mC; mA; rU; mC; rA; rG; mU; rA; zc3p$rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1738 zidB; rA; mC; rA; rA; rG;rA; rU; mU; rA; mC; rU; mC; rG; rU2p; rG; rC; rG; rA; rG; rA; mC; rG;rC; mU; rC; rG; mC; mA; rU; mC; rA; rG; mU; rA; zc3p$ rU; mU; rG; rU;zc3p; zc3p$ SERPINH1_6_S1740 zidB; rA; mC; rA; rA; rG; rA; rU; mU; rA;mC; rU; mC; rG; rU2p; rG; rC; rG; rA; rG; rA; mC; rG; rC; mU; rC; mG;rC; mA; rU; mC; rA; rG; mU; rA; zc3p$ rU; mU; rG; mU; zc3p; zc3p$SERPINH1_6_S1742 zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC;rG; mU; rC; rG; rC; rG; rA; rG; rA; mC; rG; mU; rC; mG; rC; mA; rU; mC;rA; rG; mU; rA; zc3p$ rU; mU; rG; mU; zc3p; zc3p$ SERPINH1_6_S1743 zidB;rA; rC; rA; rA; rG; rA; rU; mU; rA; mC; rU; mC; rG; mU; mC; rG; rC; rG;rA; rG; rA; mC; rG; rU; mC; rG; mC; mA; rU; mC; rA; rG; mU; rA; zc3p$rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1745 zidB; rA; rC; rA; rA; rG;rA; rU; mU; rA; mC; rU; mC; rG; rU2p; rG; rC; rG; rA; rG; rA; mC; rG;rC; mU; rC; mG; rC; mA; rU; mC; rA; rG; mU; rA; zc3p$ rU; mU; rG; rU;zc3p; zc3p$ SERPINH1_6_S1747 zidB; rA; rC; rA; rA; rG; rA; rU; mU; rA;mC; rU; mC; rG; rU2p; rG; rC; rG; rA; rG; rA; mC; rG; rC; mU; rC; rG;mC; mA; rU; rC; rA; rG; mU; rA; zc3p$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1748 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG;rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; mG; rC; mA; rU; mC;rG; rA; rG; LdT; rA$ rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1749 zidB;rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG; rU2p; mU; rG; rC; rG;rA; rG; rA; mC; rC; mU; rC; mG; rC; mA; rU; mC; rG; rA; rG; LdT; rA$ rU;mU; rG; mU; zc3p; zc3p$ SERPINH1_6_S1750 zidB; rA; mC; rA; rA; rG; rA;mU; rA; mC; rU; mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU;rC; rG; mC; mA; rU; rC; rG; rA; rG; LdT; rA$ rU; mU; rG; rU; zc3p; zc3p$SERPINH1_6_S1751 zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG;rU2p; mU; rG; rC; rG; rA; rG; rA; mC; rC; mU; rC; mG; rC; mA; rU; mC;rG; rA; rG; mU; rA; zc3p$ rU; mU; rG; rU; zc3p; zc3p$ SERPINH1_6_S1752zidB; rA; mC; rA; rA; rG; rA; mU; rA; mC; rU; mC; rG; rU2p; mU; rG; rC;rG; rA; rG; rA; mC; rC; mU; rC; mG; rC; mA; rU; mC; rG; rA; rG; mU; rA;zc3p$ rU; mU; rG; mU; zc3p; zc3p$ SERPINH1_6_S1753 zidB; rA; mC; rA; rA;rG; rA; mU; rA; mC; rU; mC; rG; rU2p; mU; rG; rC; rG; rA; rG; rA; mC;rC; mU; rC; rG; mC; mA; rU; rC; rG; rA; rG; mU; rA; zc3p$ rU; mU; rG;rU; zc3p; zc3p$ SERPINH1_42_S1354 rG; rA; rC; rA; rG; rG; rC; rC; rU;yrA; rU; rA; rG; rU; rU; rG; rU; rA; rC; rU; rA; rC; rA; rA; rC; rU; rG;rA; rG; rG; rC; rC; rU; rG; rA; yrU; zdT; zdT$ rU; rC; zdT; zdT$SERPINH1_51_S1671 zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; rA; mC; rC;rC2p; rA; mU; rA; rC; rA; mC; rA; mU; rG; rG; rU; rG; mU; mC; rU; mC;rG; rG; mU; rG; rA; zc3p$ rA; rG; rG; rA; zc3p; zc3p$ SERPINH1_51_S1672zidB; rU; rC; rC; mU; rG; rA; rG; mU; rC; rA; mC; rC; LdC; rA; mU; rA;rC; rA; mC; rA; mU; rG; rG; rU; rG; mU; mC; rU; mC; rG; rG; mU; rG; rA;zc3p$ rA; rG; rG; rA; zc3p; zc3p$ SERPINH1_51_S1681 zidB; rU; mC; rC;mU; rG; rA; mU; rC; rA; mC; rC; rC2p; rA; mU; rG; rA; rC; rA; mC; rA;mU; rG; rG; rU; rG; rU; mC; rU; mC; rA; rG; rG; mU; rG; rA; zc3p$ rG;rG; rA; zc3p; zc3p$ SERPINH1_51_S1683 zidB; rU; mC; rC; mU; rG; rA; mU;rC; mA; rC; rC; rC2p; rA; mU; rG; rA; rC; rA; mC; rA; mU; rG; rG; rU;rG; mU; mC; rU; mC; rG; rG; mU; rG; rA; zc3p$ rA; rG; rG; rA; zc3p;zc3p$

indicates data missing or illegible when filed

TABLE 7 Code of the modified nucleotides/unconventional moieties as usedin the Tables herein. Code Description rA riboadenosine-3′-phosphate;3′-adenylic acid rC ribocytidine-3′-phosphate; 3′-cytidylic acid rGriboguanosine-3′-phosphate; 3′-guanylic acid rUribouridine-3′-phosphate; 3′-uridylic acid mA2′-O-methyladenosine-3′-phosphate; 2′-O-methyl-3′-adenylic acid mC2′-O-methylcytidine-3′-phosphate; 2′-O-methyl-3′-cytidylic acid mG2′-O-methylguanosine-3′-phosphate; 2′-O-methyl-3′-guanylic acid mU2′-O-methyluridine-3′-phosphate; 2′-O-methyl-3′-uridylic acid dAdeoxyriboadenosine-3′-phosphate; 2′-deoxyribo-3′-adenylic acid dCdeoxyribocytidine-3′-phosphate; 2′-deoxyribo-3′-cytidylic acid dGdeoxyriboguanosine-3′-phosphate; 2′-deoxyribo-3′-guanylic acid dTthymidine-3′-phosphate; 3′-thymidylic acid rA2priboadenosine-2′-phosphate; 2′-adenylic acid (2′5′ A) rC2pribocytidine-2′-phosphate; 2′-cytidylic acid (2′5′ C) rG2priboguanosine-2′-phosphate; 2′-guanylic acid (2′5′ G) rU2pribouridine-2′-phosphate; 2′-uridylic acid (2′5′U) LdAL-deoxyriboadenosine-3′-phosphate (mirror image dA) LdCL-deoxyribocytidine-3′-phosphate (mirror image dC) LdGL-deoxyriboguanosine-3′-phosphate (mirror image dG) LdTL-deoxyribothymidine-3′-phosphate (mirror image dT) dB abasicdeoxyribose-3′-phosphate; 1,2-dideoxy-D-ribofuranose-3-phosphate;1,4-2-deoxy-D-ribitol-3-phosphate zidB Inverted abasicdeoxyribose-5′-phosphate; At 5′ = 5′-5′ idAb; At 3′ = 3′-3′ idAb zPrefix to indicate moiety covalently attached to 3′ terminus or 5′terminus psiU pseudouridne p 5′ phosphate s 5′ phosphorothioate C3 C3non-nucleotide $ lacking a 3′ linker (used together with abovenucleotides at the 3′ end of the sequence)

siRNA oligonucleotides useful in generating double stranded RNAmolecules are disclosed in Tables A-18, A-19 and B-E below.

SERPINH1 Oligonucleotide Sequence Useful in the Preparation of siRNACompounds.

TABLE A-18 SEQ SEQ ID ID NO NO Cross Ident Human Name SEN Sense(5′ > 3′) AS Antisense (5′ > 3′) Species gi″32454740 SERPINH1_2 60GAGACACAUGGGUGCUAUA 127 UAUAGCACCCAUGUGUCUC H, Rt, Rh, [1533-1551] M, D(18/19) SERPINH1_3 61 GGGAAGAUGCAGAAGAAGA 128 UCUUCUUCUGCAUCUUCCC H, Rt,Rh, [1112-1130] Rb (18/19) SERPINH1_5 62 GAAGAAGGCUGUUGCCAUA 129UAUGGCAACAGCCUUCUUC H, Rt [1123-1141] (18/19) SERPINH1_6 63ACAAGAUGCGAGACGAGUA 130 UACUCGUCUCGCAUCUUGU H, Rt, Rh, [1464-1482](18/19) SERPINH1_7 64 GGACAACCGUGGCUUCAUA 131 UAUGAAGCCACGGUUGUCC H, Rh,M [886-904] (18/19) SERPINH1_8 65 UGCAGUCCAUCAACGAGUA 132UACUCGUUGAUGGACUGCA H, Rt, Rh, M [738-756] (18/19) SERPINH1_9 66GCCUCAUCAUCCUCAUGCA 133 UGCAUGAGGAUGAUGAGGC H, Rt, Rh, [1026-1044] M, D(18/19) SERPINH1_10 67 CGCGCUGCAGUCCAUCAAA 134 UUUGAUGGACUGCAGCGCG H,Rt, Rh [733-751] (18/19) SERPINH1_11 68 CGGACAGGCCUCUACAACA 135UGUUGUAGAGGCCUGUCCG H, Rt, Rh, P [944-962] (18/19) SERPINH1_13 69UGACAAGAUGCGAGACGAA 136 UUCGUCUCGCAUCUUGUCA H, Rh [1462-1480] (18/19)SERPINH1_14 70 CCAGCCUCAUCAUCCUCAA 137 UUGAGGAUGAUGAGGCUGG H, M, Rt,[1023-1041] Rh, D- (18/19) SERPINH1_15 71 GCUGCAGUCCAUCAACGAA 138UUCGUUGAUGGACUGCAGC H, Rt, Rh [736-754] (18/19) SERPINH1_16 72GCAGCGCGCUGCAGUCCAA 139 UUGGACUGCAGCGCGCUGC H, Rt, Rh [729-747] (18/19)SERPINH1_17 73 UGAGACACAUGGCIUGCUAA 140 UUAGCACCCAUGUGUCUCA H, Rt, Rh[1532-1550] M, D (18/19) SERPINH1_19 74 GGUGGAGGUGACCCAUGAA 141UUCAUGGGUCACCUCCACC H, Rt, Rh, M [1159-1177] (18/19) SERPINH1_20 75CUUUGACCAGGACAUCUAA 142 UUAGAUGUCCUGGUCAAAG H, Rt, Rh [1324-1342](18/19) SERPINH1_21 76 GGAGGUGACCCAUGACCUA 143 UAGGUCAUGGGUCACCUCC H,Rt, Rh, [1162-1180] M, D (18/19) SERPINH1_22 77 CUCCUGAGACACAUGGGUA 144UACCCAUGUGUCUCAGGAG H, D [1528-1546] (18/19) SERPINH1_23 78AGAAGAAGGCUGUUGCCAA 145 UUGGCAACAGCCUUCUUCU H, Rt [1122-1140] (18/19)SERPINH1_24 79 AGCUCUCCAGCCUCAUCAA 146 UUGAUGAGGCUGGAGAGCU H, Rt, D,[1017-1035] M, P, Rh (18/19) SERPINH1_25 80 CUGCAGUCCAUCAACGAGA 147UCUCGUUGAUGGACUGCAG H, Rt, Rh M [737-755] (18/19) SERPINH1_26 81CCGGACAGGCCUCUACAAA 148 UUUGUAGAGGCCUGUCCGG H, Rt, Rh, [943-961] Rb, P(18/19) SERPINH1_27 82 GCACCGGACAGGCCUCUAA 149 UUAGAGGCCUGUCCGGUGC H,Rt, Rh, [940-958] Rb, P (18/19) SERPINH1_28 83 GCAGAAGAAGGCUGUUGCA 150UGCAACAGCCUUCUUCUGC H, Rt [1120-1138] (18/19) SERPINH1_31 84AGAAGGCUGUUGCCAUCUA 151 UAGAUGGCAACAGCCUUCU H, Rt [1125-1143] (18/19)SERPINH1_32 85 AGCGCAGCGCGCUGCAGUA 152 UACUGCAGCGCGCUGCGCU H, Rt, Rh,[726-744] (18/19) SERPINH1_33 86 GACACAUGGGUGCUAUUGA 153UCAAUAGCACCCAUGUGUC H, Rt, Rh, M [1535-1553] (18/19) SERPINH1_34 87GGGCCUGACUGAGGCCAUA 154 UAUGGCCUCAGUCAGGCCC H, Rt [1201-1219] (18/19)SERPINH1_35 88 AGACACAUGGGUGCUAUUA 155 UAAUAGCACCCAUGUGUCU H, Rt, Rh, M[1534-1552] (18/19) SERPINH1_36 89 CCAUGACCUGCAGAAACAA 156UUGUUUCUGCAGGUCAUGG H, Rt, Rh, M [1171-1189] (18/19) SERPINH1_37 90AGAUGCAGAAGAAGGCUGA 157 UCAGCCUUCUUCUGCAUCU H, Rt, Rh, M [1116-1134](18/19) SERPINH1_38 91 CAAGCUCUCCAGCCUCAUA 158 UAUGAGGCUGGAGAGCUUG H,Rt, Rh, [1015-1033] M, P, D (18/19) SERPINH1_39 92 UGCAGAAGAAGGCUGUUGA159 UCAACAGCCUUCUUCUGCA H, Rt [1119-1137] (18/19) SERPINH1_41 93CAGCCUCAUCAUCCUCAUA 160 UAUGAGGAUGAUGAGGCUG H, Rt, Rh, [1024-1042] M, D(18/19) SERPINH1_42 94 GACAGGCCUCUACAACUAA 161 UUAGUUGUAGAGGCCUGUC H,Rt, Rh, [946-964] Rb, P (18/19) SERPINH1_43 95 GAUGCAGAAGAAGGCUGUA 162UACAGCCUUCUUCUGCAUC H, Rt, Rh, M [1117-1135] (18/19) SERPINH1_44 96ACCCAUGACCUGCAGAAAA 163 UUUUCUGCAGGUCAUGGGU H, Rt, Rh, M [1169-1187](18/19) SERPINH1_45 97 ACUCCAAGAUCAACUUCCA 164 UGGAAGUUGAUCUUGGAGU H,Rt, Rh, [702-720] M, D (18/19) SERPINH1_45a 98 ACUCCAAGAUCAACUUCCU 165AGGAAGUUGAUCUUGGAGU H, Rt, Rh, [702-720] M, D (18/19) SERPINH1_48 99AGGCCUCUACAACUACUAA 166 UUAGUAGUUGUAGAGGCCU H, Rt, Rh, [949-967] Rb, P,D (18/19) SERPINH1_49 100 CACUCCAAGAUCAACUUCA 167 UGAAGUUGAUCUUGGAGUG H,Rt, Rh, [701-719] M, D (18/19) SERPINH1_51 101 UCCUGAGACACAUGGGUGA 168UCACCCAUGUGUCUCAGGA H, Rt, D, M [1529-1547] (18/19) SERPINH1_52 102GACAAGAUGCGAGACGAGA 169 UCUCGUCUCGCAUCUUGUC H, Rt, Rh, [1463-1481](18/19) SERPINH1_53 103 GGUGACCCAUGACCUGCAA 170 UUGCAGGUCAUGGGUCACC H,Rt, Rh, M [1165-1183] (18/19) SERPINH1_59 104 CCGAGGUGAAGAAACCUGA 171UCAGGUUUCUUCACCUCGG H, Rt, Rh, [285-303] (18/19) SERPINH1_51a 105UCCUGAGACACAUGGGUGU 172 ACACCCAUGUGUCUCAGGA H, Rt, D, M [1529-1547](18/19) SERPINH1_61 106 GCACUCCAAGAUCAACUUA 173 UAAGUUGAUCUUGGAGUGC H,Rh, D [700-718] (18/19) SERPINH1_62 107 GUGGUGGAGGUGACCCAUA 174UAUGGGUCACCUCCACCAC H, Rt, Rh, [1157-1175] M, Rb (18/19) SERPINH1_64 108GCCGAGGUGAAGAAACCUA 175 UAGGUUUCUUCACCUCGGC H, Rt, Rh, [284-302] (18/19)SERPINH1_65 109 GCUCUCCAGCCUCAUCAUA 176 UAUGAUGAGGCUGGAGAGC H, Rt, D,[1018-1036] M, P, Rh (18/19) SERPINH1_66 110 GAUGCACCGGACAGGCCUA 177UAGGCCUGUCCGGUGCAUC H, Rt, Rh, [937-955] M, Rb, P (18/19) SERPINH1_68111 CUCUCCAGCCUCAUCAUCA 178 UGAUGAUGAGGCUGGAGAG H, Rt, D, [1019-1037] M,P, Rh (18/19) SERPINH1_69 112 GCAGACCACCGACGGCAAA 179UUUGCCGUCGGUGGUCUGC H, Rt, D [763-781] (18/19) SERPINH1_70 113AGUCCAUCAACGAGUGGGA 180 UCCCACUCGUUGAUGGACU H, Rt, Rh, M [741-759](18/19) SERPINH1_71 114 ACCGUGGCUUCAUGGUGAA 181 UUCACCAUGAAGCCACGGU H,Rt, Rh, M [891-909] (18/19) SERPINH1_74 115 GAAGGCUGUUGCCAUCUCA 182UGAGAUGGCAACAGCCUUC H, Rt, [1126-1144] (18/19) SERPINH1_75 116GAAGAUGCAGAAGAAGGCA 183 UGCCUUCUUCUGCAUCUUC H, Rt, Rh, [1114-1132] Rb(18/19) SERPINH1_77 117 UGAUGAUGCACCGGACAGA 184 UCUGUCCGGUGCAUCAUCA H,Rh, [933-951] (18/19) SERPINH1_78 118 CCCUUUGACCAGGACAUCA 185UGAUGUCCUGGUCAAAGGG H, Rt, Rh, [1322-1340] (18/19) SERPINH1_80 119CAGUCCAUCAACGAGUGGA 186 UCCACUCGUUGAUGGACUG H, Rt, Rh, M [740-758](18/19) SERPINH1_82 120 CAACCGUGGCUUCAUGGUA 187 UACCAUGAAGCCACGGUUG H,Rt, Rh, M [889-907] (18/19) SERPINH1_83 121 CGACAAGCGCAGCGCGCUA 188UAGCGCGCUGCGCUUGUCG H [721-739] (18/19) SERPINH1_84 122GCAGUCCAUCAACGAGUGA 189 UCACUCGUUGAUGGACUGC H, Rt, Rh, M [739-757](18/19) SERPINH1_86 123 ACAGGCCUCUACAACUACA 190 UGUAGUUGUAGAGGCCUGU H,Rt, Rh, [947-965] Rb, P, D (18/19) SERPINH1_87 124 AAGAUGCAGAAGAAGGCUA191 UAGCCUUCUUCUGCAUCUU H, Rt, Rh, M [1115-1133] (18/19) SERPINH1_89 125CAGCGCGCUGCAGUCCAUA 192 UAUGGACUGCAGCGCGCUG H, Rt, Rh, [730-748] (18/19)SERPINH1_90 126 GCGCAGCGCGCUGCAGUCA 193 UGACUGCAGCGCGCUGCGC H, Rt, Rh,[727-745] (18/19) Select siRNAs SEQ ID SEQ ID Activity Activity ActivityIC50 siRNA SEN AS 0.1 nM 0.5 nM 5 nM (nM) Length SERPINH1_2 60 127 65 487 .008 19 SERPINH1_6 63 130 164 39 5 .019 19 SERPINH1_11 68 135 119 54 6.05 19 SERPINH1_13 69 136 91 24 4 19 SERPINH1_45 97 164 156 38 8 .07 19SERPINH1_45a 98 165 19 SERPINH1_51 101 168 68 39 5 .05 19 SERPINH1_52102 169 149 37 9 0.06 19 SERPINH1_86 123 190 121 61 0.27 19 SEQ SEQ IDID Activity Activity Activity Activity Activity Activity Activity siRNASEN AS 0.026 nM 0.077 nM 0.23 nM 0.69 nM 2.1 nM 6.25 nM 25 nMSERPINH1_45 97 164 102 81 55 41 28 22 16 SERPINH1_45a 98 165 107 98 18469 36 24 16

TABLE A-19 SEQ SEQ ID ID Ident NO NO Human gi Name SEN Sense (5′ > 3′)AS Antisense (5′ > 3′) Species L 32454740 SERPINH1_1 194GGACAGGCCUCUACAACUA 219 UAGUUGUAGAGGCCUGUCC H, Rt, Rh, 19 [945-963] Rb,P (19/19) SERPINH1_1 195 GAGACACAUGGGUGCUAUU 220 AAUAGCACCCAUGUGUCUC H,Rt, Rh, 19 [1533-1551] M, D (19/19) SERPINH1_12 196 ACAAGAUGCGAGACGAGUU221 AACUCGUCUCGCAUCUUGU H, Rt, Rh, 19 [1464-1482] (19/19) SERPINH1_18197 CCUUUGACCAGGACAUCUA 222 UAGAUGUCCUGGUCAAAGG H, Rt, Rh, 19[1323-1341] (19/19) SERPINH1_29 198 GACCCAUGACCUGCAGAAA 223UUUCUGCAGGUCAUGGGUC H, Rt, Rh, M 19 [1168-1186] (19/19) SERPINH1_30 199CGGACAGGCCUCUACAACU 224 AGUUGUAGAGGCCUGUCCG H, Rt, Rh, 19 [944-962] Rb,P (19/19) SERPINH1_40 200 ACCGGACAGGCCUCUACAA 225 UUGUAGAGGCCUGUCCGGU H,Rt, Rh, 19 [942-960] Rb, P, (19/19) SERPINH1_46 201 GCAGCGCGCUGCAGUCCAU226 AUGGACUGCAGCGCGCUGC H, Rt, Rh, 19 [729-747] (19/19) SERPINH1_47 202GCGCGCUGCAGUCCAUCAA 227 UUGAUGGACUGCAGCGCGC H, Rt, Rh, 19 [732-750](19/19) SERPINH1_50 203 CUGAGACACAUGGGUGCUA 228 UAGCACCCAUGUGUCUCAG H,Rt, Rh, 19 [1531-1549] M, D (19/19) SERPINH1_54 204 AGAAGAAGGCUGUUGCCAU229 AUGGCAACAGCCUUCUUCU H, Rt 19 [1122-1140] (19/19) SERPINH1_55 205AGCUCUCCAGCCUCAUCAU 230 AUGAUGAGGCUGGAGAGCU H, Rt, D, 19 [1017-1035] M,P, Rh (19/19) SERPINH1_56 206 CUGCAGUCCAUCAACGAGU 231ACUCGUUGAUGGACUGCAG H, Rt, Rh, M 19 [737-755] (19/19) SERPINH1_57 207CGCUGCAGUCCAUCAACGA 232 UCGUUGAUGGACUGCAGCG H, Rt, Rh, 19 [735-753](19/19) SERPINH1_58 208 GACAAGAUGCGAGACGAGU 233 ACUCGUCUCGCAUCUUGUC H,Rt, Rh, 19 [1463-1481] (19/19) SERPINH1_63 209 GGGCCUGACUGAGGCCAUU 234AAUGGCCUCAGUCAGGCCC H, Rt 19 [1201-1219] (19/19) SERPINH1_67 210GAUGCAGAAGAAGGCUGUU 235 AACAGCCUUCUUCUGCAUC H, Rt, Rh, M 19 [1117-1135](19/19) SERPINH1_72 211 CACCGGACAGGCCUCUACA 236 UGUAGAGGCCUGUCCGGUG H,Rt, Rh, 19 [941-959] Rb, P (19/19) SERPINH1_73 212 AGAUGCAGAAGAAGGCUGU237 ACAGCCUUCUUCUGCAUCU H, Rt, Rh M 19 [1116-1134] (19/19) SERPINH1_76713 AGCGCGCUGCAGUCCAUCA 238 UGAUGGACUGCAGCGCGCU H, Rt, Rh 19 [731-749](19/19) SERPINH1_79 214 GGAAGAUGCAGAAGAAGGC 239 GCCUUCUUCUGCAUCUUCC H,Rt, Rh, 19 [1113-1131] Rb (19/19) SERPINH1_81 215 GAAGAAGGCUGUUGCCAUC240 GAUGGCAACAGCCUUCUUC H, Rt 19 [1123-1141] (19/19) SERPINH1_85 216UGCAGUCCAUCAACGAGUG 241 CACUCGUUGAUGGACUGCA H, Rt, Rh, M 19 [738-756](19/19) SERPINH1_88 217 CCUGAGACACAUGGGUGCU 242 AGCACCCAUGUGUCUCAGG H,Rt, D, M 19 [1530-1548] (19/19) SERPINH1_91 218 CGCAGCGCGCUGCAGUCCA 243UGGACUGCAGCGCGCUGCG H, Rt, Rh, 19 [728-746] (19/19) Select siRNAs SEQ IDSEQ ID Activity Activity Activity siRNA NO SEN NO AS 0.1 nM 0.5 nM 5 nMIC50 (nM) Length SERPINH1_4 195 220  60 35 5 .006 19 SERPINH1_12 196 221 54 42 8 .065 19 SERPINH1_18 197 222 139 43 9 19 SERPINH1_30 199 224 14649 9 0.093 19 SERPINH1_58 208 233 na na 8 19 SERPINH1_88 217 242 105 439 19

TABLE B Additional Active 19-mer SERPINH1 siRNAs No SEQ ID SEN SensesiRNA SEQ ID AS AntiSense siRNA Other human- 1 244 GGCAGACUCUGGUCAAGAA460 UUCUUGACCAGAGUCUGCC Rh [2009-2027] 2 245 CAGUGAGGCGGAUUGAGAA 461UUCUCAAUCCGCCUCACUG [1967-1985] 3 246 AGCCUUUGUUGCUAUCAAU 462AUUGAUAGCAACAAAGGCU Rh [2117-2135] 4 247 CCAUGUUCUUCAAGCCACA 463UGUGGCUUGAAGAACAUGG Rh, Rb, D [837-855] 5 248 CCCUCUUCUGACACUAAAA 464UUUUAGUGUCAGAAGAGGG [1850-1868] 6 249 CCUCAAUCAGUAUUCAUAU 465AUAUGAAUACUGAUUGAGG [1774-1792] 7 250 GAGACACAUGGGUGCUAUU 466AAUAGCACCCAUGUGUCUC Rh, D, Rt, M [1533-1551] 8 251 GUGACAAGAUGCGAGACGA467 UCGUCUCGCAUCUUGUCAC Rh [1461-1479] 9 252 GCCACACUGGGAUGAGAAA 468UUUCUCAUCCCAGUGUGGC Rh, Rb, M [850-868] 10 253 AGAUGCGAGACGAGUUAUA 469UAUAACUCGUCUCGCAUCU Rh [1467-1485] 11 254 ACGACGACGAGAAGGAAAA 470UUUUCCUUCUCGUCGUCGU [966-984] 12 255 GCCUCUACAACUACUACGA 471UCGUAGUAGUUGUAGAGGC Rb, D [951-969] 13 256 AGAUCAACUUCCGCGACAA 472UUGUCGCGGAAGUUGAUCU D [708-726] 14 257 ACUACUACGACGACGAGAA 473UUCUCGUCGUCGUAGUAGU Rb [960-978] 15 258 AGCCCUCUUCUGACACUAA 474UUAGUGUCAGAAGAGGGCU [1848-1866] 16 259 ACAAGAUGCGAGACGAGUU 475AACUCGUCUCGCAUCUUGU Rh, Rt [1464-1482] 17 260 AGCCACACUGGGAUGAGAA 476UUCUCAUCCCAGUGUGGCU Rh, Rb, M [849-867] 18 261 AGGACCAGGCAGUGGAGAA 477UUCUCCACUGCCUGGUCCU Rh [408-426] 19 262 CAGGCAAGAAGGACCUGUA 478UACAGGUCCUUCUUGCCUG Rh, D [1251-1269] 20 263 ACCUGUGAGACCAAAUUGA 479UCAAUUUGGUCUCACAGGU Rh [1813-1831] 21 264 CUUUGUUGCUAUCAAUCCA 480UGGAUUGAUAGCAACAAAG Rh [2120-2138] 22 265 GUGAGACCAAAUUGAGCUA 481UAGCUCAAUUuGGUCUCAC Rh [1817-1835] 23 266 CCCUGAAAGUCCCAGAUCA 482UGAUCUGGGACUUUCAGGG [1749-1767] 24 267 CCUUUGACCAGGACAUCUA 483UAGAUGUCCUGGUCAAAGG Rh, Rt [1323-1341] 25 268 GACCAGGCAGUGGAGAACA 484UGUUCUCCACUGCCUGGUC Rh [410-428] 26 269 CGCGCAACGUGACCUGGAA 485UUCCAGGUCACGUUGCGCG M [597-615] 27 270 AUGAGAAAUUCCACCACAA 486UUGUGGUGGAAUUUCUCAU Rh [861-879] 28 271 GAAGAAACCUGCAGCCGCA 487UGCGGCUGCAGGUUUCUUC [292-310] 29 272 CUCUCGAGCGCCUUGAAAA 488UUUUCAAGGCGCUCGAGAG [1059-1077] 30 273 GGAACAUGAGCCUUUGUUG 489CAACAAAGGCUCAUGUUCC Rh [2109-2127] 31 274 CUCACCUGUGAGACCAAAU 490AUUUGGUCUCACAGGUGAG Rh [1810-1828] 32 275 CUACGACGACGAGAAGGAA 491UUCCUUCUCGUCGUCGUAG Rb [964-982] 33 276 ACCACAAGAUGGUGGACAA 492UUGUCCACCAUCUUGUGGU Rh, Rb, M, P [873-891] 34 277 CUGGCACUGCGGAGAAGUU493 AACUUCUCCGCAGUGCCAG [318-336] 35 278 GGUCCUAUACCGUGGGUGU 494ACACCCACGGUAUAGGACC Rh [912-930] 36 279 CCAACCUCUCCCAACUAUA 495UAUAGUUGGGAGAGGUUGG Rh [1896-1914] 37 280 GAGAAGGAAAAGCUGCAAA 496UUUGCAGCUUUUCCUUCUC Rh [974-992] 38 281 GCCUCUCGAGCGCCUUGAA 497UUCAAGGCGCUCGAGAGGC [1057-1075] 39 282 AGGCCAUUGACAAGAACAA 498UUGUUCUUGUCAAUGGCCU Rh, D [1212-1230] 40 283 GACCCAUGACCUGCAGAAA 499UUUCUGCAGGUCAUGGGUC Rh, Rt, M [1168-1186] 41 284 CUCCUGGCACUGCGGAGAA 500UUCUCCGCAGUGCCAGGAG [315-333] 42 285 CGGACAGGCCUCUACAACU 501AGUUGUAGAGGCCUGUCCG Rh, Rb, Rt, P [944-962] 43 286 GAUGAGAAAUUCCACCACA502 UGUGGUGGAAUUUCUCAUC Rh [860-878] 44 287 CACGCAUGUCAGGCAAGAA 503UUCUUGCCUGACAUGCGUG Rh, D [1242-1260] 45 288 ACCUCUCCCAACUAUAAAA 504UUUUAUAGUUGGGAGAGGU Rh [1899-1917] 46 289 ACCAGGCAGUGGAGAACAU 505AUGUUCUCCACUGCCUGGU Rh [411-429] 47 290 GGGAACAUGAGCCUUUGUU 506AACAAAGGCUCAUGUUCCC Rh [2108-2126] 48 291 AGAAUUCACUCCACUUGGA 507UCCAAGUGGAGUGAAUUCU Rh [1653-1671] 49 292 GGGCAGACUCUGGUCAAGA 508UCUUGACCAGAGUCUGCCC Rh [2008-2026] 50 293 AGAAGGAAAAGCUGCAAAU 509AUUUGCAGCUUUUCCUUCU Rh [975-993] 51 294 GGCAGUGGAGAACAUCCUG 510CAGGAUGUUCUCCACUGCC Rh [415-433] 52 295 GGGAUGAGAAAUUCCACCA 511UGGUGGAAUUUCUCAUCCC Rh [858-876] 53 296 CCAAGCUGUUCUACGCCGA 512UCGGCGUAGAACAGCUUGG Rh [1365-1383] 54 297 ACCGGACAGGCCUCUACAA 513UUGUAGAGGCCUGUCCGGU Rh, Rb, Rt, P [942-960] 55 298 CUGCCUCAAUCAGUAUUCA514 UGAAUACUGAUUGAGGCAG [1771-1789] 56 299 CAGCCCUCUUCUGACACUA 515UAGUGUCAGAAGAGGGCUG [1847-1865] 57 300 CCAGCCUCAUCAUCCUCAU 516AUGAGGAUGAUGAGGCUGG Rh, D, Rt, M [1023-1041] 58 301 AGGGUGACAAGAUGCGAGA517 UCUCGCAUCUUGUCACCCU Rh, D [1458-1476] 59 302 GGACCAGGCAGUGGAGAAC 518GUUCUCCACUGCCUGGUCC Rh [409-427] 60 303 GCAGCGCGCUGCAGUCCAU 519AUGGACUGCAGCGCGCUGC Rh, Rt [729-747] 61 304 GCGCGCUGCAGUCCAUCAA 520UUGAUGGACUGCAGCGCGC Rh, Rt [732-750] 62 305 CCAGAUACCAUGAUGCUGA 521UCAGCAUCAUGGUAUCUGG Rh [1680-1698] 63 306 CUAGUGCGGGACACCCAAA 522UUUGGGUGUCCCGCACUAG [1400-1418] 64 307 AGGCAGUGGAGAACAUCCU 523AGGAUGUUCUCCACUGCCU Rh [414-432] 65 308 CUGAGACACAUGGGUGCUA 524UAGCACCCAUGUGUCUCAG Rh, D, Rt, M [1531-1549] 66 309 GAUUGAGAAGGAGCUCCCA525 UGGGAGCUCCUUCUCAAUC [1977-1995] 67 310 CGCAGACCACCGACGGCAA 526UUGCCGUCGOUGGUCUGCG D, Rt [762-780] 68 311 CCACACUGGGAUGAGAAAU 527AUUUCUCAUCCCAGUGUGG Rh [851-869] 69 312 GCUCAGUGAGCUUCGCUGA 528UCAGCGAAGCUCACUGAGC [642-660] 70 313 CGCCUUUGAGUUGGACACA 529UGUGUCCAACUCAAAGGCG Rh [1294-1312] 71 314 GGGUCAGCCAGCCCUCUUC 530GAAGAGGGCUGGCUGACCC Rh [1839-1857] 72 315 GGGCUUCUGGGCAGACUCU 531AGAGUCUGCCCAGAAGCCC Rh [2000-2018] 73 316 GGUACCUUCUCACCUGUGA 532UCACAGGUGAGAAGGUACC Rh [1802-1820] 74 317 GCCUGCCUCAAUCAGUAUU 533AAUACUGAUUGAGGCAGGC [1769-1787] 75 318 UCUACAACUACUACGACGA 534UCGUCGUAGUAGUUGUAGA Rb [954-972] 76 319 GGGAAGAUGCAGAAGAAGG 535CCUUCUUCUGCAUCUUCCC Rh, Rb, Rt [1112-1130] 77 320 CGAGAAGGAAAAGCUGCAA536 UUGCAGCUUUUCCUUCUCG Rh [973-991] 78 321 AGAAGAAGGCUGUUGCCAU 537AUGGCAACAGCCUUCUUCU Rt [1122-1140] 79 322 CACAAGCUCUCCAGCCUCA 538UGAGGCUGGAGAGCUUGUG Rh, D, M, P [1013-1031] 80 323 GGGUGACAAGAUGCGAGAC539 GUCUCGCAUCUUGUCACCC Rh, D [1459-1477] 81 324 UGUUGGAGCGUGGAAAAAA 540UUUUUUCCACGCUCCAACA [2190-2208] 82 325 CUUUGAGUUGGACACAGAU 541AUCUGUGUCCAACUCAAAG Rh [1297-1315] 83 326 AGCUCUCCAGCCUCAUCAU 542AUGAUGAGGCUGGAGAGCU Rh, D, Rt, M, [1017-1035] 84 327 AGCUGUUCUACGCCGACCA543 UGGUCGGCGUAGAACAGCU Rh [1368-1386] 85 328 CUGCAGUCCAUCAACGAGU 544ACUCGUUGAUGGACUGCAG Rh, Rt, M [737-755] 86 329 UACGACGACGAGAAGGAAA 545UUUCCUUCUCGUCGUCGUA [965-983] 87 330 CCUAGUGCGGGACACCCAA 546UUGGGUGUCCCGCACUAGG [1399-1417] 88 331 CUUCUCACCUGUGAGACCA 547UGGUCUCACAGGUGAGAAG Rh [1807-1825] 89 332 AGUUGGACACAGAUGGCAA 548UUGCCAUCUGUGUCCAACU [1302-1320] 90 333 CAGUGGAGAACAUCCUGGU 549ACCAGGAUGUUCUCCACUG Rh [417-435] 91 334 CCAGCUAGAAUUCACUCCA 550UGGAGUGAAUUCUAGCUGG Rh [1647-1665] 92 335 CGCUGCAGUCCAUCAACGA 551UCGUUGAUGGACUGCAGCG Rh, Rt [735-753] 93 336 CCAAGGACCAGGCAGUGGA 552UCCACUGCCUGGUCCUUGG Rh [405-423] 94 337 AGUUCUUCAAAGAUAGGGA 553UCCCUAUCUUUGAAGAACU [2082-2100] 95 338 CGGACCUUCCCAGCUAGAA 554UUCUAGCUGGGAAGGUCCG Rh [1638-1656] 96 339 GACAAGAUGCGAGACGAGU 555ACUCGUCUCGCAUCUUGUC Rh, Rt [1463-1481] 97 340 CCAAGAUCAACUUCCGCGA 556UCGCGGAAGUUGAUCUUGG D [705-723] 98 341 CCCAUCACGUGGAGCCUCU 557AGAGGCUCCACGUGAUGGG Rh [1044-1062] 99 342 CCAUGAUGCUGAGCCCGGA 558UCCGGGCUCAGCAUCAUGG [1687-1705] 100 343 AGCCUGCCUCAAUCAGUAU 559AUACUGAUUGAGGCAGGCU [1768-1786] 101 344 CGGCCUAAGGGUGACAAGA 560UCUUGUCACCCUUAGGCCG Rh [1451-1469] 102 345 GGGCCUGACUGAGGCCAUU 561AAUGGCCUCAGUCAGGCCC Rt [1201-1219] 103 346 UCACCUGUGAGACCAAAUU 562AAUUUGGUCUCACAGGUGA Rh [1811-1829] 104 347 GAGGCCAUUGACAAGAACA 563UGUUCUUGUCAAUGGCCUC Rh, D [1211-1229] 105 348 GCUCCUGGCACUGCGGAGA 564UCUCCGCAGUGCCAGGAGC [314-332] 106 349 GGCGCCUGGUCCGGCCUAA 565UUAGGCCGGACCAGGCGCC Rh [1440-1458] 107 350 CCAGCCCUCUUCUGACACU 566AGUGUCAGAAGAGGGCUGG [1846-1864] 108 351 ACUACGACGACGAGAAGGA 567UCCUUCUCGUCGUCGUAGU Rb [963-981] 109 352 CCUAUACCGUGGGUGUCAU 568AUGACACCCACGGUAUAGG Rh, D, P [915-933] 110 353 GACCCAGCUCAGUGAGCUU 569AAGCUCACUGAGCUGGGUC [636-654] 111 354 UGGGUGUCAUGAUGAUGCA 570UGCAUCAUCAUGACACCCA Rh [924-942] 112 355 CCAAGGGUGUGGUGGAGGU 571ACCUCCACCACACCCUUGG Rh, D [1149-1167] 113 356 AGGUCACCAAGGACGUGGA 572UCCACGUCCUUGGUGACCU Rh, D [789-807] 114 357 CCCUGGCCGCCGAGGUGAA 573UUCACCUCGGCGGCCAGGG [276-294] 115 358 AGCACUCCAAGAUCAACUU 574AAGUUGAUCUUGGAGUGCU Rh, D [699-717] 116 359 CCUGGCACUGCGGAGAAGU 575ACUUCUCCGCAGUGCCAGG [317-335] 117 360 GAUGCAGAAGAAGGCUGUU 576AACAGCCUUCUUCUGCAUC Rh, Rt, M [1117-1135] 118 361 CCCACAAGCUCUCCAGCCU577 AGGCUGGAGAGCUUGUGGG Rh, D, P [1011-1029] 119 362 CUCUUCUGACACUAAAACA578 UGUUUUAGUGUCAGAAGAG [1852-1870] 120 363 ACGAGAAGGAAAAGCUGCA 579UGCAGCUUUUCCUUCUCGU Rh [972-990] 121 364 UGAAAAGCUGCUAACCAAA 580UUUGGUUAGCAGCUUUUCA [1072-1090] 122 365 UCUCACCUGUGAGACCAAA 581UUUGGUCUCACAGGUGAGA Rh [1809-1827] 123 366 CAUGAUGAUGCACCGGACA 582UGUCCGGUGCAUCAUCAUG Rh [931-949] 124 367 GGAUUGAGAAGGAGCUCCC 583GGGAGCUCCUUCUCAAUCC [1976-1994] 125 368 CCUUCAUCUUCCUAGUGCG 584CGCACUAGGAAGAUGAAGG [1389-1407] 126 369 GGCCUGGCCUUCAGCUUGU 585ACAAGCUGAAGGCCAGGCC [374-392] 127 370 GGUCAGCCAGCCCUCUUCU 586AGAAGAGGGCUGGCUGACC Rh [1840-1858] 128 371 UUCUCACCUGUGAGACCAA 587UUGGUCUCACAGGUGAGAA Rh [1808-1826] 129 372 CGCAGCAGCUCCUGGCACU 588AGUGCCAGGAGCUGCUGCG [307-325] 130 373 GCCAUGUUCUUCAAGCCAC 589GUGGCUUGAAGAACAUGGC Rh, Rb, D [836-854] 131 374 AGGCAGUGCUGAGCGCCGA 590UCGGCGCUCAGCACUGCCU [510-528] 132 375 CACCUGUGAGACCAAAUUG 591CAAUUUGGUCUCACAGGUG Rh [1812-1830] 133 376 CACCGGACAGGCCUCUACA 592UGUAGAGGCCUGUCCGGUG Rh, Rb, Rt, P [941-959] 134 377 AGCUAGAAUUCACUCCACU593 AGUGGAGUGAAUUCUAGCU Rh [1649-1667] 135 378 AGAUGCAGAAGAAGGCUGU 594ACAGCCUUCUUCUGCAUCU Rh, Rt, M [1116-1134] 136 379 CCCUGCUAGUCAACGCCAU595 AUGGCGUUGACUAGCAGGG Rh [822-840] 137 380 ACAACUACUACGACGACGA 596UCGUCGUCGUAGUAGUUGU Rb [957-975] 138 381 GCUCCUGAGACACAUGGGU 597ACCCAUGUGUCUCAGGAGC D [1527-1545] 139 382 UGGAGAACAUCCUGGUGUC 598GACACCAGGAUGUUCUCCA Rh [420-438] 140 383 AGCGCGCUGCAGUCCAUCA 599UGAUGGACUGCAGCGCGCU Rh, Rt [731-749] 141 384 CGCCUUGAAAAGCUGCUAA 600UUAGCAGCUUUUCAAGGCG [1067-1085] 142 385 GCCUUUGUUGCUAUCAAUC 601GAUUGAUAGCAACAAAGGC Rh [2118-2136] 143 386 CUCUACAACUACUACGACG 602CGUCGUAGUAGUUGUAGAG Rb [953-971] 144 387 CGCUCACUCAGCAACUCCA 603UGGAGUUGCUGAGUGAGCG Rh [575-593] 145 388 GGUACCAGCCUUGGAUACU 604AGUAUCCAAGGCUGGUACC Rh [1571-1589] 146 389 GCCUGACUGAGGCCAUUGA 605UCAAUGGCCUCAGUCAGGC Rh [1203-1221] 147 390 UGAGCUUCGCUGAUGACUU 606AAGUCAUCAGCGAAGCUCA Rh [648-666] 148 391 CCAGCCUUGGAUACUCCAU 607AUGGAGUAUCCAAGGCUGG Rh [1575-1593] 149 392 AAAGGCUCCUGAGACACAU 608AUGUGUCUCAGGAGCCUUU [1523-1541] 150 393 UGACCCAUGACCUGCAGAA 609UUCUGCAGGUCAUGGGUCA Rh, Rt, M [1167-1185] 151 394 CCUGUGAGACCAAAUUGAG610 CUCAAUUUGGUCUCACAGG Rh [1814-1832] 152 395 GCGGACCUUCCCAGCUAGA 611UCUAGCUGGGAAGGUCCGC Rh [1637-1655] 153 396 GGAAGAUGCAGAAGAAGGC 612GCCUUCUUCUGCAUCUUCC Rh, Rb, Rt [1113-1131] 154 397 UGCCCAAGGGUGUGGUGGA613 UCCACCACACCCUUGGGCA Rh, D [1146-1164] 155 398 GGAGCCUCUCGAGCGCCUU614 AAGGCGCUCGAGAGGCUCC [1054-1072] 156 399 GACUCUGGUCAAGAAGCAU 615AUGCUUCUUGACCAGAGUC Rh [2013-2031] 157 400 CAGGCAGUGGAGAACAUCC 616GGAUGUUCUCCACUGCCUG Rh [413-431] 158 401 CAAGCCUGCCUCAAUCAGU 617ACUGAUUGAGGCAGGCUUG Rh [1766-1784] 159 402 CUGGAAGCUGGGCAGCCGA 618UCGGCUGCCCAGCUUCCAG [610-628] 160 403 GAAGAAGGCUGUUGCCAUC 619GAUGGCAACAGCCUUCUUC Rt [1123-1141] 161 404 GGGCGAGCUGCUGCGCUCA 620UGAGCGCAGCAGCUCGCCC Rh [562-580] 162 405 AAGCCACACUGGGAUGAGA 621UCUCAUCCCAGUGUGGCUU Rh, Rb, M [848-866] 163 406 GUGUGGUGGAGGUGACCCA 622UGGGUCACCUCCACCACAC Rh, D [1155-1173] 164 407 CCGCCUUUGAGUUGGACAC 623GUGUCCAACUCAAAGGCGG Rh [1293-1311] 165 408 GGCCAUUGACAAGAACAAG 624CUUGUUCUUGUCAAUGGCC Rh, D [1213-1231] 166 409 UGCCUCAAUCAGUAUUCAU 625AUGAAUACUGAUUGAGGCA [1772-1790] 167 410 CCUUCCCAGCUAGAAUUCA 626UGAAUUCUAGCUGGGAAGG Rh [1642-1660] 168 411 GGGACCUGGGCCAUAGUCA 627UGACUAUGGCCCAGGUCCC [1721-1739] 169 412 CGAGGUGAAGAAACCUGCA 628UGCAGGUUUCUUCACCUCG Rh [286-304] 170 413 GCCUUUGAGUUGGACACAG 629CUGUGUCCAACUCAAAGGC Rh [1295-1313] 171 414 AGCGGACCUUCCCAGCUAG 630CUAGCUGGGAAGGUCCGCU Rh [1636-1654] 172 415 CGCAUGUCAGGCAAGAAGG 631CCUUCUUGCCUGACAUGCG Rh, D [1244-1262] 173 416 ACAACUGCGAGCACUCCAA 632UUGGAGUGCUCGCAGUUGU Rh, D [690-708] 174 417 GAGGCGGAUUGAGAAGGAG 633CUCCUUCUCAAUCCGCCUC [1971-1989] 175 418 GGCCGCCGAGGUGAAGAAA 634UUUCUUCACCUCGGCGGCC [280-298] 176 419 CAGCUCUAUCCCAACCUCU 635AGAGGUUGGGAUAGAGCUG [1886-1904] 177 420 AGCUGGGCAGCCGACUGUA 636UACAGUCGGCUGCCCAGCU [615-633] 178 421 GCCAUUGACAAGAACAAGG 637CCUUGUUCUUGUCAAUGGC Rh, D [1214-1232] 179 422 CGCCAUGUUCUUCAAGCCA 638UGGCUUGAAGAACAUGGCG Rh, Rb, P [835-853] 180 423 CCGAGGUCACCAAGGACGU 639ACGUCCUUGGUGACCUCGG Rh, D [786-804] 181 424 GGACCCAGCUCAGUGAGCU 640AGCUCACUGAGCUGGGUCC [635-653] 182 425 CCAAUGACAUUUUGUUGGA 641UCCAACAAAAUGUCAUUGG [2178-2196] 183 426 AGUGAGGCGGAUUGAGAAG 642CUUCUCAAUCCGCCUCACU [1968-1986] 184 427 UGCAGUCCAUCAACGAGUG 643CACUCGUUGAUGGACUGCA Rh, Rt, M [738-756] 185 428 UGUCACGCAUGUCAGGCAA 644UUGCCUGACAUGCGUGACA Rh, D [1239-1257] 186 429 CGACGACGAGAAGGAAAAG 645CUUUUCCUUCUCGUCGUCG [967-985] 187 430 ACAAGAACAAGGCCGACUU 646AAGUCGGCCUUGUUCUUGU Rh [1221-1239] 188 431 CUUCAAGCCACACUGGGAU 647AUCCCAGUGUGGCUUGAAG Rh, Rb, D [844-862] 189 432 CCUGGGCCAUAGUCAUUCU 648AGAAUGACUAUGGCCCAGG [1725-1743] 190 433 UUUGUUGGAGCGUGGAAAA 649UUUUCCACGCUCCAACAAA [2188-2206] 191 434 AGAACAUCCUGGUGUCACC 650GGUGACACCAGGAUGUUCU [423-441] 192 435 ACGCCACCGCCUUUGAGUU 651AACUCAAAGGCGGUGGCGU Rh [1287-1305] 193 436 GUGAGGUACCAGCCUUGGA 652UCCAAGGCUGGUACCUCAC Rh [1567-1585] 194 437 GCGCCUUCUGCCUCCUGGA 653UCCAGGAGGCAGAAGGCGC [252-270] 195 438 GCCUGGCCUUCAGCUUGUA 654UACAAGCUGAAGGCCAGGC [375-393] 196 439 CCCGGAAACUCCACAUCCU 655AGGAUGUGGAGUUUCCGGG [1700-1718] 197 440 UCUUCAAGCCACACUGGGA 656UCCCAGUGUGGCUUGAAGA Rh, Rb, D [843-861] 198 441 UGUUGCUAUCAAUCCAAGA 657UCUUGGAUUGAUAGCAACA Rh [2123-2141] 199 442 GAGUGGGCCGCGCAGACCA 658UGGUCUGCGCGGCCCACUC [752-770] 200 443 CCUGAGACACAUGGGUGCU 659AGCACCCAUGUGUCUCAGG D, Rt, M [1530-1548] 201 444 AGCCGACUGUACGGACCCA 660UGGGUCCGUACAGUCGGCU [623-641] 202 445 GGGCCUCAGGGUGCACACA 661UGUGUGCACCCUGAGGCCC [1486-1504] 203 446 ACUGGGAUGAGAAAUUCCA 662UGGAAUUUCUCAUCCCAGU Rh [855-873] 204 447 AGAAUGACCUGGCCGCAGU 663ACUGCGGCCAGGUCAUUCU [1952-1970] 205 448 CAUAUUUAUAGCCAGGUAC 664GUACCUGGCUAUAAAUAUG Rh [1788-1806] 206 449 AGGUGACCCAUGACCUGCA 665UGCAGGUCAUGGGUCACCU Rh, Rt, M [1164-1182] 207 450 GCGCUGCAGUCCAUCAACG666 CGUUGAUGGACUGCAGCGC Rh, Rt [734-752] 208 451 GGUGACAAGAUGCGAGACG 667CGUCUCGCAUCUUGUCACC Rh [1460-1478] 209 452 CUUCAAAGAUAGGGAGGGA 668UCCCUCCCUAUCUUUGAAG [2086-2104] 210 453 AGCUGCAAAUCGUGGAGAU 669AUCUCCACGAUUUGCAGCU Rh [984-1002] 211 454 GUGGAGAACAUCCUGGUGU 670ACACCAGGAUGUUCUCCAC Rh [419-437] 212 455 GAACAAGGCCGACUUGUCA 671UGACAAGUCGGCCUUGUUC Rh [1225-1243] 213 456 CAUGAUGCUGAGCCCGGAA 672UUCCGGGCUCAGCAUCAUG [1688-1706] 214 457 GCGCCUUGAAAAGCUGCUA 673UAGCAGCUUUUCAAGGCGC Rh [1066-1084] 215 458 GCAGACUCUGGUCAAGAAG 674CUUCUUGACCAGAGUCUGC Rh [2010-2028] 216 459 CCAGGCAGUGGAGAACAUC 675GAUGUUCUCCACUGCCUGG Rh [412-430]

TABLE C Cross-Species 19-mer SERPINH1 siRNAs No. SEQ ID SEN Sense siRNASEQ ID AS AntiSense siRNA Other Species human- 1 676 CACUACAACUGCGAGCACU973 AGUGCUCGCAGUUGUAGUG Rh, D [686-704] 2 677 AACCGUGGCUUCAUGGUGA 974UCACCAUGAAGCCACGGUU Rh, Rt, M [890-908] 3 678 GGCAAGAAGGACCUGUACC 975GGUACAGGUCCUUCUUGCC Rh, D, M [1253-      4 679 GGUGGACAACCGUGGCUUC 976GAAGCCACGGUUGUCCACC Rh, M [883-901] 5 680 AGGCCAUGGCCAAGGACCA 977UGGUCCUUGGCCAUGGCCU Rh, D [396-414] 6 681 CGCAGCGCGCUGCAGUCCA 978UGGACUGCAGCGCGCUGCG Rh, Rt [728-746] 7 682 AGCAGCAAGCAGCACUACA 979UGUAGUGCUGCUUGCUGCU Rh, D [674-692] 8 683 GGCCUCUACAACUACUACG 980CGUAGUAGUUGUAGAGGCC Rb, D [950-968] 9 684 GAAGAUGCAGAAGAAGGCU 981AGCCUUCUUCUGCAUCUUC Rh, Rb, Rt [1114-      10 685 GGCUCCUGAGACACAUGGG982 CCCAUGUGUCUCAGGAGCC D [1526-      11 686 AGCAAGCAGCACUACAACU 983AGUUGUAGUGCUGCUUGCU Rh, D [677-695] 12 687 GGAGGUGACCCAUGACCUG 984CAGGUCAUGGGUCACCUCC Rh, Rt, M [1162-      13 688 CCCUUUGACCAGGACAUCU 985AGAUGUCCUGGUCAAAGGG Rh, Rt [1322-      14 689 CUCCUGAGACACAUGGGUG 986CACCCAUGUGUCUCAGGAG D [1528-      15 690 AAGGCUCCUGAGACACAUG 987CAUGUGUCUCAGGAGCCUU D [1524-      16 691 CGCGCUGCAGUCCAUCAAC 988GUUGAUGGACUGCAGCGCG Rh, Rt [733-751] 17 692 AGGGUGUGGUGGAGGUGAC 989GUCACCUCCACCACACCCU Rh, D [1152-      18 693 AGCACUACAACUGCGAGCA 990UGCUCGCAGUUGUAGUGCU Rh, D [684-702] 19 694 GGCUCCCUGCUAUUCAUUG 991CAAUGAAUAGCAGGGAGCC D [1421-      20 695 GCGCGCAACGUGACCUGGA 992UCCAGGUCACGUUGCGCGC M [596-614] 21 696 GCUGCAGUCCAUCAACGAG 993CUCGUUGAUGGACUGCAGC Rh, Rt [736-754] 22 697 ACCAAAGAGCAGCUGAAGA 994UCUUCAGCUGCUCUUUGGU Rh, Rb, P [1085-      23 698 CCAAGGACGUGGAGCGCAC 995GUGCGCUCCACGUCCUUGG Rh, D [795-813] 24 699 UGUUCUUCAAGCCACACUG 996CAGUGUGGCUUGAAGAACA Rh, Rb, D [840-858] 25 700 GCCCAAGGGUGUGGUGGAG 997CUCCACCACACCCUUGGGC Rh, D [1147-      26 701 ACAGGCCUCUACAACUACU 998AGUAGUUGUAGAGGCCUGU Rh, Rb, D, Rt, P [947-965] 27 702UGCGCAGCAGCAAGCAGCA 999 UGCUGCUUGCUGCUGCGCA Rh, D [669-687] 28 703GGUGGAGGUGACCCAUGAC 1000 GUCAUGGGUCACCUCCACC Rh, Rt, M [1159-      29704 CUUUGACCAGGACAUCUAC 1001 GUAGAUGUCCUGGUCAAAG Rh, Rt [1324-      30705 AAGGGUGUGGUGGAGGUGA 1002 UCACCUCCACCACACCCUU Rh, D [1151-      31706 UCCUAUACCGUGGGUGUCA 1003 UGACACCCACGGUAUAGGA Rh, D, P [914-932] 32707 GCGCAGACCACCGACGGCA 1004 UGCCGUCGGUGGUCUGCGC D [761-779] 33 708CGCAGCAGCAAGCAGCACU 1005 AGUGCUGCUUGCUGCUGCG Rh, D [671-689] 34 709GCCUCAUCAUCCUCAUGCC 1006 GGCAUGAGGAUGAUGAGGC Rh, D, Rt, M [1026-      35710 UCUCCAGCCUCAUCAUCCU 1007 AGGAUGAUGAGGCUGGAGA Rh, D, Rt, M[1020-      36 711 CCAUUGACAAGAACAAGGC 1008 GCCUUGUUCUUGUCAAUGG Rh, D[1215-      37 712 AGCAGCACUACAACUGCGA 1009 UCGCAGUUGUAGUGCUGCU Rh, D[681-699] 38 713 UGCACCGGACAGGCCUCUA 1010 UAGAGGCCUGUCCGGUGCA Rh, Rb,Rt, P [939-957] 39 714 ACUCCAAGAUCAACUUCCG 1011 CGGAAGUUGAUCUUGGAGU Rh,D, Rt, M [702-720] 40 715 UGGACAACCGUGGCUUCAU 1012 AUGAAGCCACGGUUGUCCARh, M [885-903] 41 716 GAGCAGCUGAAGAUCUGGA 1013 UCCAGAUCUUCAGCUGCUC Rh,D [1091-      42 717 CAGAAGAAGGCUGUUGCCA 1014 UGGCAACAGCCUUCUUCUG Rt[1121-      43 718 AGGCAAGAAGGACCUGUAC 1015 GUACAGGUCCUUCUUGCCU Rh, D[1252-      44 719 CCUCUACAACUACUACGAC 1016 GUCGUAGUAGUUGUAGAGG Rb, D[952-970] 45 720 AGCAGCUGAAGAUCUGGAU 1017 AUCCAGAUCUUCAGCUGCU Rh, D[1092-      46 721 AACUACUACGACGACGAGA 1018 UCUCGUCGUCGUAGUAGUU Rb[959-977] 47 722 GGCAAGCUGCCCGAGGUCA 1019 UGACCUCGGGCAGCUUGCC Rh, D[776-794] 48 723 CCGGACAGGCCUCUACAAC 1020 GUUGUAGAGGCCUGUCCGG Rh, Rb,Rt, P [943-961] 49 724 GCUCCCUGCUAUUCAUUGG 1021 CCAAUGAAUAGCAGGGAGC D[1422-      50 725 AACUGCGAGCACUCCAAGA 1022 UCUUGGAGUGCUCGCAGUU Rh, D[692-710] 51 726 GACACAUGGGUGCUAUUGG 1023 CCAAUAGCACCCAUGUGUC Rh, Rt, M[1535-      52 727 GCACCGGACAGGCCUCUAC 1024 GUAGAGGCCUGUCCGGUGC Rh, Rb,Rt, P [940-958] 53 728 AGCGCAGCGCGCUGCAGUC 1025 GACUGCAGCGCGCUGCGCU Rh,Rt [726-744] 54 729 GGACGUGGAGCGCACGGAC 1026 GUCCGUGCGCUCCACGUCC Rh, D[799-817] 55 730 CAGCCUCAUCAUCCUCAUG 1027 CAUGAGGAUGAUGAGGCUG Rh, D, Rt,M [1024-      56 731 AAGAUCAACUUCCGCGACA 1028 UGUCGCGGAAGUUGAUCUU D[707-725] 57 732 GCGCAACGUGACCUGGAAG 1029 CUUCCAGGUCACGUUGCGC M[598-616] 58 733 ACUGCGAGCACUCCAAGAU 1030 AUCUUGGAGUGCUCGCAGU Rh, D[693-711] 59 734 GUGGACAACCGUGGCUUCA 1031 UGAAGCCACGGUUGUCCAC Rh, M[884-902] 60 735 CCACAAGCUCUCCAGCCUC 1032 GAGGCUGGAGAGCUUGUGG Rh, D, P[1012-      61 736 CAAGAUGGUGGACAACCGU 1033 ACGGUUGUCCACCAUCUUG Rh, Rb,M, P [877-895] 62 737 CGAGCACUCCAAGAUCAAC 1034 GUUGAUCUUGGAGUGCUCG Rh, D[697-715] 63 738 AGCUGCCCGAGGUCACCAA 1035 UUGGUGACCUCGGGCAGCU Rh, D[780-798] 64 739 GGACAUCUACGGGCGCGAG 1036 CUCGCGCCCGUAGAUGUCC D[1333-      65 740 AGGACAUCUACGGGCGCGA 1037 UCGCGCCCGUAGAUGUCCU D[1332-      66 741 UGUCAGGCAAGAAGGACCU 1038 AGGUCCUUCUUGCCUGACA Rh, D[1248-      67 742 GGGUGUGGUGGAGGUGACC 1039 GGUCACCUCCACCACACCC Rh, D[1153-      68 743 CAAGCUCUCCAGCCUCAUC 1040 GAUGAGGCUGGAGAGCUUG Rh, D,M, P [1015-      69 744 GUGACCCAUGACCUGCAGA 1041 UCUGCAGGUCAUGGGUCAC Rh,Rt, M [1166-      70 745 GUUCUUCAAGCCACACUGG 1042 CCAGUGUGGCUUGAAGAACRh, Rb, D [841-859] 71 746 ACAUCUACGGGCGCGAGGA 1043 UCCUCGCGCCCGUAGAUGUD, M [1335-      72 747 UGGAGGUGACCCAUGACCU 1044 AGGUCAUGGGUCACCUCCA Rh,Rt, M [1161-      73 748 UGCAGAAGAAGGCUGUUGC 1045 GCAACAGCCUUCUUCUGCA Rt[1119-      74 749 UGUACCAGGCCAUGGCCAA 1046 UUGGCCAUGGCCUGGUACA Rh, D[390-408] 75 750 UGUGGUGGAGGUGACCCAU 1047 AUGGGUCACCUCCACCACA Rh, D[1156-      76 751 AGAAGGACCUGUACCUGGC 1048 GCCAGGUACAGGUCCUUCU Rh, D[1257-      77 752 AGCAGCUGCGCGACGAGGA 1049 UCCUCGUCGCGCAGCUGCU Rh, D[528-546] 78 753 ACGCCAUGUUCUUCAAGCC 1050 GGCUUGAAGAACAUGGCGU Rh, Rb, P[834-852] 79 754 ACAAGAUGGUGGACAACCG 1051 CGGUUGUCCACCAUCUUGU Rh, Rb, M,P [876-894] 80 755 CUGCGAGCACUCCAAGAUC 1052 GAUCUUGGAGUGCUCGCAG Rh, D[694-712] 81 756 GUCACGCAUGUCAGGCAAG 1053 CUUGCCUGACAUGCGUGAC Rh, D[1240-      82 757 ACGCAUGUCAGGCAAGAAG 1054 CUUCUUGCCUGACAUGCGU Rh, D[1243-      83 758 UGCUAUUCAUUGGGCGCCU 1055 AGGCGCCCAAUGAAUAGCA D[1428-      84 759 UGCGCGACGAGGAGGUGCA 1056 UGCACCUCCUCGUCGCGCA Rh, D[534-552] 85 760 GCAGCUGAAGAUCUGGAUG 1057 CAUCCAGAUCUUCAGCUGC Rh, D[1093-      86 761 CCAUGACCUGCAGAAACAC 1058 GUGUUUCUGCAGGUCAUGG Rh, Rt,M [1171-      87 762 AAGCUCUCCAGCCUCAUCA 1059 UGAUGAGGCUGGAGAGCUU Rh, D,Rt, M, P [1016-      88 763 CAGCAAGCAGCACUACAAC 1060 GUUGUAGUGCUGCUUGCUGRh, D [676-694] 89 764 AUGUUCUUCAAGCCACACU 1061 AGUGUGGCUUGAAGAACAU Rh,Rb, D [839-857] 90 765 UCCUGAGACACAUGGGUGC 1062 GCACCCAUGUGUCUCAGGA D,Rt, M [1529-      91 766 CACUCCAAGAUCAACUUCC 1063 GGAAGUUGAUCUUGGAGUGRh, D, Rt, M [701-719] 92 767 AAGGGUGACAAGAUGCGAG 1064CUCGCAUCUUGUCACCCUU Rh, D [1457-      93 768 GACAGGCCUCUACAACUAC 1065GUAGUUGUAGAGGCCUGUC Rh, Rb, Rt, P [946-964] 94 769 ACCCAUGACCUGCAGAAAC1066 GUUUCUGCAGGUCAUGGGU Rh, Rt, M [1169-      95 770CACCACAAGAUGGUGGACA 1067 UGUCCACCAUCUUGUGGUG Rh, Rb, M, P [872-890] 96771 GCAGAAGAAGGCUGUUGCC 1068 GGCAACAGCCUUCUUCUGC Rt [1120-      97 772GUGGUGGAGGUGACCCAUG 1069 CAUGGGUCACCUCCACCAC Rh, Rb, Rt, M [1157-     98 773 AGGCCUCUACAACUACUAC 1070 GUAGUAGUUGUAGAGGCCU Rh, Rb, D, Rt, P[949-967] 99 774 GGUGACCCAUGACCUGCAG 1071 CUGCAGGUCAUGGGUCACC Rh, Rt, M[1165-      100 775 GCCGAGGUGAAGAAACCUG 1072 CAGGUUUCUUCACCUCGGC Rh, Rt[284-302] 101 776 CAACUACUACGACGACGAG 1073 CUCGUCGUCGUAGUAGUUG Rb[958-976] 102 777 CAAGAAGGACCUGUACCUG 1074 CAGGUACAGGUCCUUCUUG Rh, D, M[1255-      103 778 UGUUCCACGCCACCGCCUU 1075 AAGGCGGUGGCGUGGAACA D[1281-      104 779 CCCUGCUAUUCAUUGGGCG 1076 CGCCCAAUGAAUAGCAGGG D[1425-      105 780 CCGUGGCUUCAUGGUGACU 1077 AGUCACCAUGAAGCCACGG Rh, Rt,M [892-910] 106 781 CUACAACUACUACGACGAC 1078 GUCGUCGUAGUAGUUGUAG Rb[955-973] 107 782 GCAGCACUACAACUGCGAG 1079 CUCGCAGUUGUAGUGCUGC Rh, D[682-700] 108 783 UGGUGGACAACCGUGGCUU 1080 AAGCCACGGUUGUCCACCA Rh, M[882-900] 109 784 AGACCACCGACGGCAAGCU 1081 AGCUUGCCGUCGGUGGUCU D, Rt[765-783] 110 785 AGAAACACCUGGCUGGGCU 1082 AGCCCAGCCAGGUGUUUCU D[1182-      111 786 ACCAAGGACGUGGAGCGCA 1083 UGCGCUCCACGUCCUUGGU Rh, D[794-812] 112 787 CCGAGGUGAAGAAACCUGC 1084 GCAGGUUUCUUCACCUCGG Rh, Rt[285-303] 113 788 ACUACAACUGCGAGCACUC 1085 GAGUGCUCGCAGUUGUAGU Rh, D[687-705] 114 789 ACAAGCUCUCCAGCCUCAU 1086 AUGAGGCUGGAGAGCUUGU Rh, D, M,P [1014-      115 790 AGGACGUGGAGCGCACGGA 1087 UCCGUGCGCUCCACGUCCU Rh, D[798-816] 116 791 GCUAUUCAUUGGGCGCCUG 1088 CAGGCGCCCAAUGAAUAGC D[1429-      117 792 AACUUCCGCGACAAGCGCA 1089 UGCGCUUGUCGCGGAAGUU D[713-731] 118 793 GCUCUCCAGCCUCAUCAUC 1090 GAUGAUGAGGCUGGAGAGC Rh, D,Rt, M, P [1018-      119 794 AGAAGGCUGUUGCCAUCUC 1091GAGAUGGCAACAGCCUUCU Rt [1125-      120 795 GGUCACCAAGGACGUGGAG 1092CUCCACGUCCUUGGUGACC Rh, D [790-808] 121 796 AGCUGCGCGACGAGGAGGU 1093ACCUCCUCGUCGCGCAGCU Rh, D [531-549] 122 797 CCCGAGGUCACCAAGGACG 1094CGUCCUUGGUGACCUCGGG Rh, D [785-803] 123 798 AUGUCAGGCAAGAAGGACC 1095GGUCCUUCUUGCCUGACAU Rh, D [1247-      124 799 CGAGGUCACCAAGGACGUG 1096CACGUCCUUGGUGACCUCG Rh, D [787-805] 125 800 GAUGCACCGGACAGGCCUC 1097GAGGCCUGUCCGGUGCAUC Rh, Rb, Rt, M, P [937-955] 126 801GCACUACAACUGCGAGCAC 1098 GUGCUCGCAGUUGUAGUGC Rh, D [685-703] 127 802CCACAAGAUGGUGGACAAC 1099 GUUGUCCACCAUCUUGUGG Rh, Rb, M, P [874-892] 128803 CAAGGGUGUGGUGGAGGUG 1100 CACCUCCACCACACCCUUG Rh, D [1150-      129804 AGCUGAAGAUCUGGAUGGG 1101 CCCAUCCAGAUCUUCAGCU Rh, D [1095-      130805 ACCAGGCCAUGGCCAAGGA 1102 UCCUUGGCCAUGGCCUGGU Rh, D [393-411] 131 806CAUGUUCUUCAAGCCACAC 1103 GUGUGGCUUGAAGAACAUG Rh, Rb, D [838-856] 132 807CAAGAUCAACUUCCGCGAC 1104 GUCGCGGAAGUUGAUCUUG D [706-724] 133 808UCCAGCCUCAUCAUCCUCA 1105 UGAGGAUGAUGAGGCUGGA Rh, D, Rt, M [1022-     134 809 GCCCGAGGUCACCAAGGAC 1106 GUCCUUGGUGACCUCGGGC Rh, D [784-802] 135810 UCAAGCCACACUGGGAUGA 1107 UCAUCCCAGUGUGGCUUGA Rh, Rb [846-864] 136811 AGUCCAUCAACGAGUGGGC 1108 GCCCACUCGUUGAUGGACU Rh, Rt, M [741-759] 137812 GACUUCGUGCGCAGCAGCA 1109 UGCUGCUGCGCACGAAGUC Rh, D, M [662-680] 138813 CUCUCCAGCCUCAUCAUCC 1110 GGAUGAUGAGGCUGGAGAG Rh, D, Rt, M, P[1019-      139 814 GCAGACCACCGACGGCAAG 1111 CUUGCCGUCGGUGGUCUGC D, Rt[763-781] 140 815 AUGCAGAAGAAGGCUGUUG 1112 CAACAGCCUUCUUCUGCAU Rt[1118-      141 816 CAACCGUGGCUUCAUGGUG 1113 CACCAUGAAGCCACGGUUG Rh, Rt,M [889-907] 142 817 UACUACGACGACGAGAAGG 1114 CCUUCUCGUCGUCGUAGUA Rb[962-980] 143 818 GAAGGCUGUUGCCAUCUCC 1115 GGAGAUGGCAACAGCCUUC Rt[1126-      144 819 UCACCAAGGACGUGGAGCG 1116 CGCUCCACGUCCUUGGUGA Rh, D[792-810] 145 820 CAGCUGAAGAUCUGGAUGG 1117 CCAUCCAGAUCUUCAGCUG Rh, D[1094-      146 821 UGGGCCUGACUGAGGCCAU 1118 AUGGCCUCAGUCAGGCCCA Rt[1200-      147 822 ACCGUGGCUUCAUGGUGAC 1119 GUCACCAUGAAGCCACGGU Rh, Rt,M [891-909] 148 823 CAGUCCAUCAACGAGUGGG 1120 CCCACUCGUUGAUGGACUG Rh, Rt,M [740-758] 149 824 CCGACGGCAAGCUGCCCGA 1121 UCGGGCAGCUUGCCGUCGG D[771-789] 150 825 ACAAGCGCAGCGCGCUGCA 1122 UGCAGCGCGCUGCGCUUGU Rh, Rt[723-741] 151 826 GAAACACCUGGCUGGGCUG 1123 CAGCCCAGCCAGGUGUUUC D[1183-      152 827 AGGCUCCUGAGACACAUGG 1124 CCAUGUGUCUCAGGAGCCU D[1525-      153 828 CAAGGACGUGGAGCGCACG 1125 CGUGCGCUCCACGUCCUUG Rh, D[796-814] 154 829 GCAGUCCAUCAACGAGUGG 1126 CCACUCGUUGAUGGACUGC Rh, Rt, M[739-757] 155 830 AGAUGGUGGACAACCGUGG 1127 CCACGGUUGUCCACCAUCU Rh, M[879-897] 156 831 AAGCGCAGCGCGCUGCAGU 1128 ACUGCAGCGCGCUGCGCUU Rh, Rt[725-743] 157 832 CAUGUCAGGCAAGAAGGAC 1129 GUCCUUCUUGCCUGACAUG Rh, D[1246-      158 833 CAAGCCACACUGGGAUGAG 1130 CUCAUCCCAGUGUGGCUUG Rh, Rb[847-865] 159 834 AAGAUGCAGAAGAAGGCUG 1131 CAGCCUUCUUCUGCAUCUU Rh, Rt, M[1115-      160 835 GGCCAUGGCCAAGGACCAG 1132 CUGGUCCUUGGCCAUGGCC Rh, D[397-415] 161 836 GUGCGCAGCAGCAAGCAGC 1133 GCUGCUUGCUGCUGCGCAC Rh, D[668-686] 162 837 CAACUGCGAGCACUCCAAG 1134 CUUGGAGUGCUCGCAGUUG Rh, D[691-709] 163 838 UACAACUGCGAGCACUCCA 1135 UGGAGUGCUCGCAGUUGUA Rh, D[689-707] 164 839 CAUUGACAAGAACAAGGCC 1136 GGCCUUGUUCUUGUCAAUG Rh, D[1216-      165 840 CAAGCAGCACUACAACUGC 1137 GCAGUUGUAGUGCUGCUUG Rh, D[679-697] 166 841 GUGUUCCACGCCACCGCCU 1138 AGGCGGUGGCGUGGAACAC D[1280-      167 842 CCUGCUAUUCAUUGGGCGC 1139 GCGCCCAAUGAAUAGCAGG D[1426-      168 843 GCCCACAAGCUCUCCAGCC 1140 GGCUGGAGAGCUUGUGGGC Rh, D,P [1010-      169 844 CAGCAGCAAGCAGCACUAC 1141 GUAGUGCUGCUUGCUGCUG Rh, D[673-691] 170 845 UGAUGCACCGGACAGGCCU 1142 AGGCCUGUCCGGUGCAUCA Rh, Rb,Rt, M, P [936-954] 171 846 UCAACUUCCGCGACAAGCG 1143 CGCUUGUCGCGGAAGUUGAD [711-729] 172 847 UCAGGCAAGAAGGACCUGU 1144 ACAGGUCCUUCUUGCCUGA Rh, D[1250-      173 848 ACUUCGUGCGCAGCAGCAA 1145 UUGCUGCUGCGCACGAAGU Rh, D,M [663-681] 174 849 ACAACCGUGGCUUCAUGGU 1146 ACCAUGAAGCCACGGUUGU Rh, Rt,M [888-906] 175 850 AAGGCUGUUGCCAUCUCCU 1147 AGGAGAUGGCAACAGCCUU D, Rt[1127-      176 851 GCAGCUGCGCGAGGAGGAG 1148 CUCCUCGUCGCGCAGCUGC Rh, D[529-547] 177 852 UAUUCAUUGGGCGCCUGGU 1149 ACCAGGCGCCCAAUGAAUA D[1431-      178 853 UCCACCACAAGAUGGUGGA 1150 UCCACCAUCUUGUGGUGGA Rh, Rb,D, P [870-888] 179 854 CCCUGGCCCACAAGCUCUC 1151 GAGAGCUUGUGGGCCAGGG Rh,D, P [1005-      180 855 ACCAGGACAUCUACGGGCG 1152 CGCCCGUAGAUGUCCUGGU D,Rt [1329-      181 856 GAUGAUGCACCGGACAGGC 1153 GCCUGUCCGGUGCAUCAUC Rh,Rb, Rt, M [934-952] 182 857 CAACGCCAUGUUCUUCAAG 1154 CUUGAAGAACAUGGCGUUGRh, Rb, P [832-850] 183 858 ACGGCAAGCUGCCCGAGGU 1155 ACCUCGGGCAGCUUGCCGURh, D [774-792] 184 859 CAGCGCGCUGCAGUCCAUC 1156 GAUGGACUGCAGCGCGCUG Rh,Rt [730-748] 185 860 CCCAAGGGUGUGGUGGAGG 1157 CCUCCACCACACCCUUGGG Rh, D[1148-      186 861 CAUGGCCAAGGACCAGGCA 1158 UGCCUGGUCCUUGGCCAUG Rh, D[400-418] 187 862 CUCCAGCCUCAUCAUCCUC 1159 GAGGAUGAUGAGGCUGGAG Rh, D,Rt, M [1021-      188 863 UCUACGGGCGCGAGGAGCU 1160 AGCUCCUCGCGCCCGUAGAD, M [1338-      189 864 GGCCCACAAGCUCUCCAGC 1161 GCUGGAGAGCUUGUGGGCCRh, D, P [1009-      190 865 GUCAGGCAAGAAGGACCUG 1162CAGGUCCUUCUUGCCUGAC Rh, D [1249-      191 866 CAUCUACGGGCGCGAGGAG 1163CUCCUCGCGCCCGUAGAUG D, M [1336-      192 867 CGUGCGCAGCAGCAAGCAG 1164CUGCUUGCUGCUGCGCACG Rh, D, M [667-685] 193 868 AGCCUCAUCAUCCUCAUGC 1165GCAUGAGGAUGAUGAGGCU Rh, D, Rt, M [1025-      194 869 UUCAAGCCACACUGGGAUG1166 CAUCCCAGUGUGGCUUGAA Rh, Rb [845-863] 195 870 AAGAAGGCUGUUGCCAUCU1167 AGAUGGCAACAGCCUUCUU Rt [1124-      196 871 GGUGUGGUGGAGGUGACCC 1168GGGUCACCUCCACCACACC Rh, D [1154-      197 872 GAGGUGACCCAUGACCUGC 1169GCAGGUCAUGGGUCACCUC Rh, Rt, M [1163-      198 873 GUGGAGGUGACCCAUGACC1170 GGUCAUGGGUCACCUCCAC Rh, Rt, M [1160-      199 874CACAAGAUGGUGGACAACC 1171 GGUUGUCCACCAUCUUGUG Rh, Rb, M, P [875-893] 200875 CUGGCCCACAAGCUCUCCA 1172 UGGAGAGCUUGUGGGCCAG Rh, D, P [1007-     201 876 GAUGACUUCGUGCGCAGCA 1173 UGCUGCGCACGAAGUCAUC Rh, Rt, M [659-677]202 877 ACUUCCGCGACAAGCGCAG 1174 CUGCGCUUGUCGCGGAAGU D [714-732] 203 878AACGCCAUGUUCUUCAAGC 1175 GCUUGAAGAACAUGGCGUU Rh, Rb, P [833-851] 204 879GGACCUGUACCUGGCCAGC 1176 GCUGGCCAGGUACAGGUCC Rh, D [1261-      205 880GCGACGAGGAGGUGCACGC 1177 GCGUGCACCUCCUCGUCGC D [537-555] 206 881GCAAGCUGCCCGAGGUCAC 1178 GUGACCUCGGGCAGCUUGC Rh, D [777-795] 207 882AUUCAUUGGGCGCCUGGUC 1179 GACCAGGCGCCCAAUGAAU D [1432-      208 883GAGGUCACCAAGGACGUGG 1180 CCACGUCCUUGGUGACCUC Rh, D [788-806] 209 884AAGAAGGACCUGUACCUGG 1181 CCAGGUACAGGUCCUUCUU Rh, D [1256-      210 885GACAACCGUGGCUUCAUGG 1182 CCAUGAAGCCACGGUUGUC Rh, Rt, M [887-905] 211 886CUGGGCCUGACUGAGGCCA 1183 UGGCCUCAGUCAGGCCCAG Rt [1199-      212 887CUCCAAGAUCAACUUCCGC 1184 GCGGAAGUUGAUCUUGGAG Rh, D, Rt, M [703-721] 213888 CAACUUCCGCGACAAGCGC 1185 GCGCUUGUCGCGGAAGUUG D [712-730] 214 889CUCCCUGCUAUUCAUUGGG 1186 CCCAAUGAAUAGCAGGGAG D [1423-      215 890AAGCAGCACUACAACUGCG 1187 CGCAGUUGUAGUGCUGCUU Rh, D [680-698] 216 891GCGCAGCAGCAAGCAGCAC 1188 GUGCUGCUUGCUGCUGCGC Rh, D [670-688] 217 892CAGGCCAUGGCCAAGGACC 1189 GGUCCUUGGCCAUGGCCUG Rh, D [395-413] 218 893GUACCAGGCCAUGGCCAAG 1190 CUUGGCCAUGGCCUGGUAC Rh, D [391-409] 219 894CUUCGUGCGCAGCAGCAAG 1191 CUUGCUGCUGCGCACGAAG Rh, D, M [664-682] 220 895CAGCACUACAACUGCGAGC 1192 GCUCGCAGUUGUAGUGCUG Rh, D [683-701] 221 896UACAACUACUACGACGACG 1193 CGUCGUCGUAGUAGUUGUA Rb [956-974] 222 897GAUGGUGGACAACCGUGGC 1194 GCCACGGUUGUCCACCAUC Rh, M [880-898] 223 898CUACAACUGCGAGCACUCC 1195 GGAGUGCUCGCAGUUGUAG Rh, D [688-706] 224 899AAGGACCUGUACCUGGCCA 1196 UGGCCAGGUACAGGUCCUU Rh, D [1259-      225 900GCUGCCCGAGGUCACCAAG 1197 CUUGGUGACCUCGGGCAGC Rh, D [781-799] 226 901GACAUCUACGGGCGCGAGG 1198 CCUCGCGCCCGUAGAUGUC D, M [1334-      227 902CCACCACAAGAUGGUGGAC 1199 GUCCACCAUCUUGUGGUGG Rh, Rb, D, P [871-889] 228903 GCGCGACGAGGAGGUGCAC 1200 GUGCACCUCCUCGUCGCGC Rh, D [535-553] 229 904CUAUUCAUUGGGCGCCUGG 1201 CCAGGCGCCCAAUGAAUAG D [1430-      230 905CCAGGACAUCUACGGGCGC 1202 GCGCCCGUAGAUGUCCUGG D, Rt [1330-      231 906AAGAUGGUGGACAACCGUG 1203 CACGGUUGUCCACCAUCUU Rh, M [878-896] 232 907CAGGACAUCUACGGGCGCG 1204 CGCGCCCGUAGAUGUCCUG D [1331-      233 908UCCAAGAUCAACUUCCGCG 1205 CGCGGAAGUUGAUCUUGGA D [704-722] 234 909GUCACCAAGGACGUGGAGC 1206 GCUCCACGUCCUUGGUGAC Rh, D [791-809] 235 910CUGCCCGAGGUCACCAAGG 1207 CCUUGGUGACCUCGGGCAG Rh, D [782-800] 236 911GACCAGGACAUCUACGGGC 1208 GCCCGUAGAUGUCCUGGUC D, Rt [1328-      237 912CCAUGGCCAAGGACCAGGC 1209 GCCUGGUCCUUGGCCAUGG Rh, D [399-417] 238 913CACCAAGGACGUGGAGCGC 1210 GCGCUCCACGUCCUUGGUG Rh, D [793-811] 239 914GACAAGCGCAGCGCGCUGC 1211 GCAGCGCGCUGCGCUUGUC Rh, Rt [722-740] 240 915CAAGCGCAGCGCGCUGCAG 1212 CUGCAGCGCGCUGCGCUUG Rh, Rt [724-742] 241 916CAGACCACCGACGGCAAGC 1213 GCUUGCCGUCGGUGGUCUG D, Rt [764-782] 242 917GACCACCGACGGCAAGCUG 1214 CAGCUUGCCGUCGGUGGUC D, Rt [766-784] 243 918AGGACCUGUACCUGGCCAG 1215 CUGGCCAGGUACAGGUCCU Rh, D [1260-      244 919CUGCUAUUCAUUGGGCGCC 1216 GGCGCCCAAUGAAUAGCAG D [1427-      245 920UCAUUGGGCGCCUGGUCCG 1217 CGGACCAGGCGCCCAAUGA Rh, D [1434-      246 921GCUGCGCGACGAGGAGGUG 1218 CACCUCCUCGUCGCGCAGC Rh, D [532-550] 247 922CGGCAAGCUGCCCGAGGUC 1219 GACCUCGGGCAGCUUGCCG Rh, D [775-793] 248 923CCUCAUCAUCCUCAUGCCC 1220 GGGCAUGAGGAUGAUGAGG Rh, D, Rt, M [1027-     249 924 CCAGGCCAUGGCCAAGGAC 1221 GUCCUUGGCCAUGGCCUGG Rh, D [394-412] 250925 GCCAUGGCCAAGGACCAGG 1222 CCUGGUCCUUGGCCAUGGC Rh, D [398-416] 251 926CCACCGACGGCAAGCUGCC 1223 GGCAGCUUGCCGUCGGUGG D, Rt [768-786] 252 927AUGGUGGACAACCGUGGCU 1224 AGCCACGGUUGUCCACCAU Rh, M [881-899] 253 928CUUCCGCGACAAGCGCAGC 1225 GCUGCGCUUGUCGCGGAAG D [715-733] 254 929CGCGACGAGGAGGUGCACG 1226 CGUGCACCUCCUCGUCGCG Rh, D [536-554] 255 930UGGCCCACAAGCUCUCCAG 1227 CUGGAGAGCUUGUGGGCCA Rh, D, P [1008-      256931 GAGCAGCUGCGCGACGAGG 1228 CCUCGUCGCGCAGCUGCUC Rh, D [527-545] 257 932UGACCAGGACAUCUACGGG 1229 CCCGUAGAUGUCCUGGUCA Rt [1327-      258 913ACCACCGACGGCAAGCUGC 1230 GCAGCUUGCCGUCGGUGGU D, Rt [767-785] 259 934GAAGGACCUGUACCUGGCC 1231 GGCCAGGUACAGGUCCUUC Rh, D [1258-      260 935CAUUGGGCGCCUGGUCCGG 1232 CCGGACCAGGCGCCCAAUG Rh, D [1435-      261 936AUGCACCGGACAGGCCUCU 1233 AGAGGCCUGUCCGGUGCAU Rh, Rb, Rt, P [938-956] 262937 AUCAACUUCCGCGACAAGC 1234 GCUUGUCGCGGAAGUUGAU D [710-728] 263 938CAGCUGCGCGACGAGGAGG 1235 CCUCCUCGUCGCGCAGCUG Rh, D [530-548] 264 939CAGAAACACCUGGCUGGGC 1236 GCCCAGCCAGGUGUUUCUG D [1181-      265 940CUACGGGCGCGAGGAGCUG 1237 CAGCUCCUCGCGCCCGUAG D, M [1339-      266 941CGACGAGGAGGUGCACGCC 1238 GGCGUGCACCUCCUCGUCG D [538-556] 267 942UUUGACCAGGACAUCUACG 1239 CGUAGAUGUCCUGGUCAAA Rt [1325-      268 943GUCCAUCAACGAGUGGGCC 1240 GGCCCACUCGUUGAUGGAC Rh, Rt, M [742-760] 269 944AUGACUUCGUGCGCAGCAG 1241 CUGCUGCGCACGAAGUCAU Rh, Rt, M [660-678] 270 945UCCCUGCUAUUCAUUGGGC 1242 GCCCAAUGAAUAGCAGGGA D [1424-      271 946CUGCGCGACGAGGAGGUGC 1243 GCACCUCCUCGUCGCGCAG Rh, D [533-551] 272 947CAAGCUGCCCGAGGUCACC 1244 GGUGACCUCGGGCAGCUUG Rh, D [778-796] 273 948AAGCUGCCCGAGGUCACCA 1245 UGGUGACCUCGGGCAGCUU Rh, D [779-797] 274 949UUCUUCAAGCCACACUGGG 1246 CCCAGUGUGGCUUGAAGAA Rh, Rb, D [842-860] 275 950ACACCUGGCUGGGCUGGGC 1247 GCCCAGCCCAGCCAGGUGU D [1186-      276 951UCCAUCAACGAGUGGGCCG 1248 CGGCCCACUCGUUGAUGGA Rt, M [743-761] 277 952AUCUACGGGCGCGAGGAGC 1249 GCUCCUCGCGCCCGUAGAU D, M [1337-      278 953UCGUGCGCAGCAGCAAGCA 1250 UGCUUGCUGCUGCGCACGA Rh, D, M [666-684] 279 954CGACGGCAAGCUGCCCGAG 1251 CUCGGGCAGCUUGCCGUCG D [772-790] 280 955UUCAUUGGGCGCCUGGUCC 1252 GGACCAGGCGCCCAAUGAA Rh, D [1433-      281 956UUGACCAGGACAUCUACGG 1253 CCGUAGAUGUCCUGGUCAA Rt [1326-      282 957CCUGGCCCACAAGCUCUCC 1254 GGAGAGCUUGUGGGCCAGG Rh, D, P [1006-      283958 UGACUUCGUGCGCAGCAGC 1255 GCUGCUGCGCACGAAGUCA Rh, Rt, M [661-679] 284959 AUGAUGCACCGGACAGGCC 1256 GGCCUGUCCGGUGCAUCAU Rh, Rb, Rt, M, P[935-953] 285 960 CACCGACGGCAAGCUGCCC 1257 GGGCAGCUUGCCGUCGGUG D, Rt[769-787] 286 961 GACGGCAAGCUGCCCGAGG 1258 CCUCGGGCAGCUUGCCGUC Rh, D[773-791] 287 962 UACCAGGCCAUGGCCAAGG 1259 CCUUGGCCAUGGCCUGGUA Rh, D[392-410] 288 963 UCCGCGACAAGCGCAGCGC 1260 GCGCUGCGCUUGUCGCGGA D[717-735] 289 964 UUCCGCGACAAGCGCAGCG 1261 CGCUGCGCUUGUCGCGGAA D[716-734] 290 965 AAGGACGUGGAGCGCACGG 1262 CCGUGCGCUCCACGUCCUU Rh, D[797-815] 291 966 UUCCACCACAAGAUGGUGG 1263 CCACCAUCUUGUGGUGGAA Rh, Rb,D, P [869-887] 292 967 UACGGGCGCGAGGAGCUGC 1264 GCAGCUCCUCGCGCCCGUA D, M[1340-      293 968 AAACACCUGGCUGGGCUGG 1265 CCAGCCCAGCCAGGUGUUU D[1184-      294 969 AACACCUGGCUGGGCUGGG 1266 CCCAGCCCAGCCAGGUGUU D[1185-      295 970 AUUGGGCGCCUGGUCCGGC 1267 GCCGGACCAGGCGCCCAAU Rh, D[1436-      296 971 ACCGACGGCAAGCUGCCCG 1268 CGGGCAGCUUGCCGUCGGU D[770-788] 297 972 UUCGUGCGCAGCAGCAAGC 1269 GCUUGCUGCUGCGCACGAA Rh, D, M[665-683]

TABLE D SERPINH1 Active 18 + 1-mer siRNAs No SEQ ID Sense siRNA SEQ IDAntiSense siRNA Other Sp human- 1 1270 AGCCUUUGUUGCUAUCAAA 1849UUUGAUAGCAACAAAGGCU Rh [2117-2135] 2 1271 GCCUAAGGGUGACAAGAUA 1850UAUCUUGUCACCCUUAGGC Rh [1453-1471] 3 1272 GGCCUAAGGGUGACAAGAA 1851UUCUUGUCACCCUUAGGCC Rh [1452-1470] 4 1273 CCUCAAUCAGUAUUCAUAA 1852UUAUGAAUACUGAUUGAG [1774-1792] 5 1274 GGCGGAUUGAGAAGGAGCA 1853UGCUCCUUCUCAAUCCGCC [1973-1991] 6 1275 GGCAGUGGAGAACAUCCUA 1854UAGGAUGUUCUCCACUGCC Rh [415-433] 7 1276 GGGUCAGCCAGCCCUCUUA 1855UAAGAGGGCUGGCUGACCC Rh [1839-1857] 8 1277 GGGUGACAAGAUGCGAGAA 1856UUCUCGCAUCUUGUCACCC Rh, D [1459-1477] 9 1278 GGACCAGGCAGUGGAGAAA 1857UUUCUCCACUGCCUGGUCC Rh [409-427] 10 1279 GAGACACAUGGGUGCUAUA 1858UAUAGCACCCAUGUGUCUC Rh, D, Rt, [1533-1551] 11 1280 GUUGGAGCGUGGAAAAAAA1859 UUUUUUUCCACGCUCCAAC [2191-2208] 12 1281 GGAACAUGAGCCUUUGUUA 1860UAACAAAGGCUCAUGUUCC Rh [2109-2127] 13 1282 GCCAUGUUCUUCAAGCCAA 1861UUGGCUUGAAGAACAUGGC Rh, Rb, D [836-854] 14 1283 GGAUUGAGAAGGAGCUCCA 1862UGGAGCUCCUUCUCAAUCC [1976-1994] 15 1284 GGGAUGAACUUUUUGUUUA 1863UAAACAAAAAGUUCAUCCC Rh [2048-2066] 16 1285 GCCGCAGUGAGGCGGAUUA 1864UAAUCCGCCUCACUGCGGC [1963-1981] 17 1286 GGACCUUCCCAGCUAGAAA 1865UUUCUAGCUGGGAAGGUCC Rh [1639-1657] 18 1287 GACCUUCCCAGCUAGAAUA 1866UAUUCUAGCUGGGAAGGUC Rh [1640-1658] 19 1288 CCUGUGAGACCAAAUUGAA 1867UUCAAUUUGGUCUCACAGG Rh [1814-1832] 20 1289 UGGAGAACAUCCUGGUGUA 1868UACACCAGGAUGUUCUCCA Rh [420-438] 21 1290 GCCUUUGUUGCUAUCAAUA 1869UAUUGAUAGCAACAAAGGC Rh [2118-2136] 22 1291 CCGCCUUUGAGUUGGACAA 1870UUGUCCAACUCAAAGGCGG Rh [1293-1311] 23 1292 CAGGCAGUGGAGAACAUCA 1871UGAUGUUCUCCACUGCCUG Rh [413-431] 24 1293 CACCUGUGAGACCAAAUUA 1872UAAUUUGGUCUCACAGGUG Rh [1812-1830] 25 1294 GGGAAGAUGCAGAAGAAGA 1873UCUUCUUCUGCAUCUUCCC Rh, Rb, Rt [1112-1130] 26 1295 GGCCAUUGACAAGAACAAA1874 UUUGUUCUUGUCAAUGGCC Rh, D [1213-1231] 27 1296 GCCUUUGAGUUGGACACAA1875 UUGUGUCCAACUCAAAGGC Rh [1295-1313] 28 1297 AGCGGACCUUCCCAGCUAA 1876UUAGCUGGGAAGGUCCGCU Rh [1636-1654] 29 1298 GAAGAAGGCUGUUGCCAUA 1877UAUGGCAACAGCCUUCUUC Rt [1123-1141] 30 1299 ACAAGAUGCGAGACGAGUA 1878UACUCGUCUCGCAUCUUGU Rh, Rt [1464-1482] 31 1300 GAGGCGGAUUGAGAAGGAA 1879UUCCUUCUCAAUCCGCCUC [1971-1989] 32 1301 GGACAACCGUGGCUUCAUA 1880UAUGAAGCCACGGUUGUCC Rh, M [886-904] 33 1302 CAUAUUUAUAGCCAGGUAA 1881UUACCUGGCUAUAAAUAUG Rh [1788-1806] 34 1303 CGACGACGAGAAGGAAAAA 1882UUUUUCCUUCUCGUCGUCG [967-985] 35 1304 CUCACCUGUGAGACCAAAA 1883UUUUGGUCUCACAGGUGAG Rh [1810-1828] 36 1305 GCGGCUCCCUGCUAUUCAA 1884UUGAAUAGCAGGGAGCCGC [1419-1437] 37 1306 AGAACAUCCUGGUGUCACA 1885UGUGACACCAGGAUGUUCU [423-441] 38 1307 CACACUGGGAUGAGAAAUA 1886UAUUUCUCAUCCCAGUGUG Rh [852-870] 39 1308 GCUAGAAUUCACUCCACUA 1887UAGUGGAGUGAAUUCUAGC Rh [1650-1668] 40 1309 CCUUCAUCUUCCUAGUGCA 1888UGCACUAGGAAGAUGAAGG [1389-1407] 41 1310 UGCUAUCAAUCCAAGAACA 1889UGUUCUUGGAUUGAUAGCA Rh [2126-2144] 42 1311 GGAAGAUGCAGAAGAAGGA 1890UCCUUCUUCUGCAUCUUCC Rh, Rb, Rt [1113-1131] 43 1312 CAUGAGCCUUUGUUGCUAA1891 UUAGCAACAAAGGCUCAUG Rh [2113-2131] 44 1313 GCGGAUUGAGAAGGAGCUA 1892UAGCUCCUUCUCAAUCCGC [1974-1992] 45 1314 UGCAGUCCAUCAACGAGUA 1893UACUCGUUGAUGGACUGCA Rh, Rt, M [738-756] 46 1315 GCACUGCGGAGAAGUUGAA 1894UUCAACUUCUCCGCAGUGC [321-339] 47 1316 CCAGGCAGUGGAGAACAUA 1895UAUGUUCUCCACUGCCUGG Rh [412-430] 48 1317 GGCAAGAAGGACCUGUACA 1896UGUACAGGUCCUUCUUGCC Rh, D, M [1253-1271] 49 1318 CUCUACAACUACUACGACA1897 UGUCGUAGUAGUUGUAGA Rb [953-971] 50 1319 CUUCCCAGCUAGAAUUCAA 1898UUGAAUUCUAGCUGGGAAG Rh [1643-1661] 51 1320 AGGCGGAUUGAGAAGGAGA 1899UCUCCUUCUCAAUCCGCCU [1972-1990] 52 1321 GGUCCUAUACCGUGGGUGA 1900UCACCCACGGUAUAGGACC Rh [912-930] 53 1322 GCAAGAAGGACCUGUACCA 1901UGGUACAGGUCCUUCUUGC Rh, D, M [1254-1272] 54 1323 CCGUGGGUGUCAUGAUGAA1902 UUCAUCAUGACACCCACGG Rh [921-939] 55 1324 GAUGCGAGACGAGUUAUAA 1903UUAUAACUCGUCUCGCAUC Rh [1468-1486] 56 1325 GGCAGUGCUGAGCGCCGAA 1904UUCGGCGCUCAGCACUGCC [511-529] 57 1326 CAGCUAGAAUUCACUCCAA 1905UUGGAGUGAAUUCUAGCUG Rh [1648-1666] 58 1327 GAGCUUCGCUGAUGACUUA 1906UAAGUCAUCAGCGAAGCUC Rh [649-667] 59 1328 CUUUGAGUUGGACACAGAA 1907UUCUGUGUCCAACUCAAAG Rh [1297-1315] 60 1329 GGUGGACAACCGUGGCUUA 1908UAAGCCACGGUUGUCCACC Rh, M [883-901] 61 1330 GCCUCAUCAUCCUCAUGCA 1909UGCAUGAGGAUGAUGAGGC Rh, D, Rt, [1026-1044] 62 1331 ACCAGGCAGUGGAGAACAA1910 UUGUUCUCCACUGCCUGGU Rh [411-429] 63 1332 CCUGCCUCAAUCAGUAUUA 1911UAAUACUGAUUGAGGCAGG [1770-1788] 64 1333 GAUCAAGCCUGCCUCAAUA 1912UAUUGAGGCAGGCUUGAUC Rh [1763-1781] 65 1334 CAGACUCUGGUCAAGAAGA 1913UCUUCUUGACCAGAGUCUG Rh [2011-2029] 66 1335 CGCGCUGCAGUCCAUCAAA 1914UUUGAUGGACUGCAGCGCG Rh, Rt [733-751] 67 1336 CUGGCACUGCGGAGAAGUA 1915UACUUCUCCGCAGUGCCAG [318-336] 68 1337 CCAGCUCUAUCCCAACCUA 1916UAGGUUGGGAUAGAGCUG [1885-1903] 69 1338 AGGGUGUGGUGGAGGUGAA 1917UUCACCUCCACCACACCCU Rh, D [1152-1170] 70 1339 AGUGAGGCGGAUUGAGAAA 1918UUUCUCAAUCCGCCUCACU [1968-1986] 71 1340 CGGACAGGCCUCUACAACA 1919UGUUGUAGAGGCCUGUCCG Rh, Rb, Rt, [944-962] 72 1341 CGACGAGAAGGAAAAGCUA1920 UAGCUUUUCCUUCUCGUCG Rh [970-988] 73 1342 AGGCCAAGGCAGUGCUGAA 1921UUCAGCACUGCCUUGGCCU Rh [504-522] 74 1343 GCCUCAGGGUGCACACAGA 1922UCUGUGUGCACCCUGAGGC [1488-1506] 75 1344 GGAUGAGAAAUUCCACCAA 1923UUGGUGGAAUUUCUCAUCC Rh [859-877] 76 1345 AGAAGGAAAAGCUGCAAAA 1924UUUUGCAGCUUUUCCUUCU Rh [975-993] 77 1346 AGCUCUAUCCCAACCUCUA 1925UAGAGGUUGGGAUAGAGC Rh [1887-1905] 78 1347 UGACAAGAUGCGAGACGAA 1926UUCGUCUCGCAUCUUGUCA Rh [1462-1480] 79 1348 AGAAGGAGCUCCCAGGAGA 1927UCUCCUGGGAGCUCCUUCU [1982-2000] 80 1349 CCUUCUCACCUGUGAGACA 1928UGUCUCACAGGUGAGAAGG Rh [1806-1824] 81 1350 GGCUUCUGGGCAGACUCUA 1929UAGAGUCUGCCCAGAAGCC Rh [2001-2019] 82 1351 CCAGCCUCAUCAUCCUCAA 1930UUGAGGAUGAUGAGGCUG Rh, D, Rt, [1023-1041] 83 1352 CCAAAGGCUCCUGAGACAA1931 UUGUCUCAGGAGCCUUUGG [1521-1539] 84 1353 GGACCUGGGCCAUAGUCAA 1932UUGACUAUGGCCCAGGUCC [1722-1740] 85 1354 GGGUGUCAUGAUGAUGCAA 1933UUGCAUCAUCAUGACACCC Rh [925-943] 86 1355 GUACCAGCCUUGGAUACUA 1934UAGUAUCCAAGGCUGGUAC Rh [1572-1590] 87 1356 GGCUGUUGCCAUCUCCUUA 1935UAAGGAGAUGGCAACAGCC [1129-1147] 88 1357 CGCAGUGAGGCGGAUUGAA 1936UUCAAUCCGCCUCACUGCG [1965-1983] 89 1358 CCAAGGACGUGGAGCGCAA 1937UUGCGCUCCACGUCCUUGG Rh, D [795-813] 90 1359 GGCUCCUGAGACACAUGGA 1938UCCAUGUGUCUCAGGAGCC D [1526-1544] 91 1360 GCUGCAGUCCAUCAACGAA 1939UUCGUUGAUGGACUGCAGC Rh, Rt [736-754] 92 1361 CCAGGUACCUUCUCACCUA 1940UAGGUGAGAAGGUACCUGG Rh [1799-1817] 93 1362 GCAGCGCGCUGCAGUCCAA 1941UUGGACUGCAGCGCGCUGC Rh, Rt [729-747] 94 1363 GAGACCAAAUUGAGCUAGA 1942UCUAGCUCAAUUUGGUCUC Rh [1819-1837] 95 1364 GCCGCCGAGGUGAAGAAAA 1943UUUUCUUCACCUCGGCGGC [281-299] 96 1365 GCAGACUCUGGUCAAGAAA 1944UUUCUUGACCAGAGUCUGC Rh [2010-2028] 97 1366 CUAGAAUUCACUCCACUUA 1945UAAGUGGAGUGAAUUCUA Rh [1651-1669] 98 1367 GCAGUGGAGAACAUCCUGA 1946UCAGGAUGUUCUCCACUGC Rh [416-434] 99 1368 CGCAUGUCAGGCAAGAAGA 1947UCUUCUUGCCUGACAUGCG Rh, D [1244-1262] 10 1369 CGGAUUGAGAAGGAGCUCA 1948UGAGCUCCUUCUCAAUCCG [1975-1993] 10 1370 AGGUGAGGUACCAGCCUUA 1949UAAGGCUGGUACCUCACCU Rh [1565-1583] 10 1371 CCACACUGGGAUGAGAAAA 1950UUUUCUCAUCCCAGUGUGG Rh [851-869] 10 1372 GCCAUUGACAAGAACAAGA 1951UCUUGUUCUUGUCAAUGGC Rh, D [1214-1232] 10 1373 GCGCUGCAGUCCAUCAACA 1952UGUUGAUGGACUGCAGCGC Rh, Rt [734-752] 10 1374 CUCCCAACUAUAAAACUAA 1953UUAGUUUAUAGUUGGGA Rh [1903-1921] 10 1375 GGUGACAAGAUGCGAGACA 1954UGUCUCGCAUCUUGUCACC Rh [1460-1478] 10 1376 GGCCGACUUGUCACGCAUA 1955UAUGCGUGACAAGUCGGCC Rh [1231-1249] 10 1377 CCUAAGGGUGACAAGAUGA 1956UCAUCUUGUCACCCUUAGG Rh [1454-1472] 10 1378 UGAGACACAUGGGUGCUAA 1957UUAGCACCCAUGUGUCUCA Rh, D, Rt, [1532-1550] 11 1379 GGGUGGAAAAACAGACCGA1958 UCGGUCUGUUUUUCCACCC [1601-1619] 11 1380 GGUGGAGGUGACCCAUGAA 1959UUCAUGGGUCACCUCCACC Rh, Rt, M [1159-1177] 11 1381 CUUUGACCAGGACAUCUAA1960 UUAGAUGUCCUGGUCAAAG Rh, Rt [1324-1342] 11 1382 GAACAUGAGCCUUUGUUGA1961 UCAACAAAGGCUCAUGUUC Rh [2110-2128] 11 1383 AGCCUUGGAUACUCCAUGA 1962UCAUGGAGUAUCCAAGGCU Rh [1577-1595] 11 1384 GGAGGUGACCCAUGACCUA 1963UAGGUCAUGGGUCACCUCC Rh, Rt, M [1162-1180] 11 1385 AGAUCAAGCCUGCCUCAAA1964 UUUGAGGCAGGCUUGAUCU Rh [1762-1780] 11 1386 GCCCAAGGGUGUGGUGGAA 1965UUCCACCACACCCUUGGGC Rh, D [1147-1165] 11 1387 AGAACAAGGCCGACUUGUA 1966UACAAGUCGGCCUUGUUCU Rh [1224-1242] 11 1388 GUGGCUUCAUGGUGACUCA 1967UGAGUCACCAUGAAGCCAC Rh [894-912] 12 1389 CUCCUGAGACACAUGGGUA 1968UACCCAUGUGUCUCAGGAG D [1528-1546] 12 1390 CAGCCUUGGAUACUCCAUA 1969UAUGGAGUAUCCAAGGCUG Rh [1576-1594] 12 1391 AAGGCUCCUGAGACACAUA 1970UAUGUGUCUCAGGAGCCUU D [1524-1542] 12 1392 AGAAGAAGGCUGUUGCCAA 1971UUGGCAACAGCCUUCUUCU Rt [1122-1140] 12 1393 CUACUACGACGACGAGAAA 1972UUUCUCGUCGUCGUAGUAG Rb [961-979] 12 1394 CCUUUGUUGCUAUCAAUCA 1973UGAUUGAUAGCAACAAAGG Rh [2119-2137] 12 1395 AGGCAGUGGAGAACAUCCA 1974UGGAUGUUCUCCACUGCCU Rh [414-432] 12 1396 CCAUCACGUGGAGCCUCUA 1975UAGAGGCUCCACGUGAUGG Rh [1045-1063] 12 1397 AGCUCUCCAGCCUCAUCAA 1976UUGAUGAGGCUGGAGAGCU Rh, D, Rt, [1017-1035] 12 1398 GGCUCCCUGCUAUUCAUUA1977 UAAUGAAUAGCAGGGAGCC D [1421-1439] 13 1399 GGGAACAUGAGCCUUUGUA 1978UACAAAGGCUCAUGUUCCC Rh [2108-2126] 13 1400 GGGCCAUAGUCAUUCUGCA 1979UGCAGAAUGACUAUGGCCC [1728-1746] 13 1401 CCAAAGAGCAGCUGAAGAA 1980UUCUUCAGCUGCUCUUUGG Rh, Rb, P [1086-1104] 13 1402 GACGAGAAGGAAAAGCUGA1981 UCAGCUUUUCCUUCUCGUC Rh [971-989] 13 1403 GGGCUUCUGGGCAGACUCA 1982UGAGUCUGCCCAGAAGCCC Rh [2000-2018] 13 1404 CAAGGACCAGGCAGUGGAA 1983UUCCACUGCCUGGUCCUUG Rh [406-424] 13 1405 CUGUGAGACCAAAUUGAGA 1984UCUCAAUUUGGUCUCACAG Rh [1815-1833] 13 1406 GACUGAGGCCAUUGACAAA 1985UUUGUCAAUGGCCUCAGUC Rh [1207-1225] 13 1407 GACUUGUCACGCAUGUCAA 1986UUGACAUGCGUGACAAGUC Rh [1235-1253] 13 1408 GAGGUGAGGUACCAGCCUA 1987UAGGCUGGUACCUCACCUC [1564-1582] 14 1409 CAGAUACCAUGAUGCUGAA 1988UUCAGCAUCAUGGUAUCUG Rh [1681-1699] 14 1410 AGGCAAGAAGGACCUGUAA 1989UUACAGGUCCUUCUUGCCU Rh, D [1252-1270] 14 1411 CUGGGAUGAGAAAUUCCAA 1990UUGGAAUUUCUCAUCCCAG Rh [856-874] 14 1412 AGGUACCAGCCUUGGAUAA 1991UUAUCCAAGGCUGGUACCU Rh [1570-1588] 14 1413 CAGCCAGCCCUCUUCUGAA 1992UUCAGAAGAGGGCUGGCUG [1843-1861] 14 1414 GUGUCAUGAUGAUGCACCA 1993UGGUGCAUCAUCAUGACAC Rh [927-945] 14 1415 CCUCUACAACUACUACGAA 1994UUCGUAGUAGUUGUAGAG Rb, D [952-970] 14 1416 CCGCCGAGGUGAAGAAACA 1995UGUUUCUUCACCUCGGCGG Rh [282-300] 14 1417 GCUAUCAAUCCAAGAACUA 1996UAGUUCUUGGAUUGAUAGC Rh [2127-2145] 14 1418 AGCCUGCCUCAAUCAGUAA 1997UUACUGAUUGAGGCAGGCU [1768-1786] 15 1419 GGUCCGGCCUAAGGGUGAA 1998UUCACCCUUAGGCCGGACC Rh [1447-1465] 15 1420 GAAGGAAAAGCUGCAAAUA 1999UAUUUGCAGCUUUUCCUUC Rh [976-994] 15 1421 GGCCUCUACAACUACUACA 2000UGUAGUAGUUGUAGAGGCC Rb, D [950-968] 15 1422 UGUUCUUCAAGCCACACUA 2001UAGUGUGGCUUGAAGAACA Rh, Rb, D [840-858] 15 1423 GGCCAAGGCAGUGCUGAGA 2002UCUCAGCACUGCCUUGGCC Rh [505-523] 15 1424 AGAAAUUCCACCACAAGAA 2003UUCUUGUGGUGGAAUUUCU Rh [864-882] 15 1425 CUGCAGUCCAUCAACGAGA 2004UCUCGUUGAUGGACUGCAG Rh, Rt, M [737-755] 15 1426 CCAGCGUGUUCCACGCCAA 2005UUGGCGUGGAACACGCUGG [1275-1293] 15 1427 GCUCCCUCCUGCUUCUCAA 2006UUGAGAAGCAGGAGGGAGC [234-252] 15 1428 CCGGACAGGCCUCUACAAA 2007UUUGUAGAGGCCUGUCCGG Rh, Rb, Rt, [943-961] 16 1429 CCCAUCACGUGGAGCCUCA2008 UGAGGCUCCACGUGAUGGG Rh [1044-1062] 16 1430 CCGGCCUAAGGGUGACAAA 2009UUUGUCACCCUUAGGCCGG Rh [1450-1468] 16 1431 CCUAUACCGUGGGUGUCAA 2010UUGACACCCACGGUAUAGG Rh, D, P [915-933] 16 1432 CAGUGGAGAACAUCCUGGA 2011UCCAGGAUGUUCUCCACUG Rh [417-435] 16 1433 CACUGGGAUGAGAAAUUCA 2012UGAAUUUCUCAUCCCAGUG Rh [854-872] 16 1434 AUCCAAAGGCUCCUGAGAA 2013UUCUCAGGAGCCUUUGGAU [1519-1537] 16 1435 UGAGAAAUUCCACCACAAA 2014UUUGUGGUGGAAUUUCUCA Rh [862-880] 16 1436 GGUGGAAAAACAGACCGGA 2015UCCGGUCUGUUUUUCCACC [1602-1620] 16 1437 GCUGGGCAGCCGACUGUAA 2016UUACAGUCGGCUGCCCAGC [616-634] 16 1438 CCAUAGUCAUUCUGCCUGA 2017UCAGGCAGAAUGACUAUGG [1731-1749] 17 1439 GCACCGGACAGGCCUCUAA 2018UUAGAGGCCUGUCCGGUGC Rh, Rb, Rt, [940-958] 17 1440 GUUGGACACAGAUGGCAAA2019 UUUGCCAUCUGUGUCCAAC [1303-1321] 17 1441 GCCUGCCUCAAUCAGUAUA 2020UAUACUGAUUGAGGCAGGC [1769-1787] 17 1442 GAUCAACUUCCGCGACAAA 2021UUUGUCGCGGAAGUUGAUC D [709-727] 17 1443 GGCCGCAGUGAGGCGGAUA 2022UAUCCGCCUCACUGCGGCC [1962-1980] 17 1444 CUGCGGAGAAGUUGAGCCA 2023UGGCUCAACUUCUCCGCAG [324-342] 17 1445 GCAUCCAAAGGCUCCUGAA 2024UUCAGGAGCCUUUGGAUGC [1517-1535] 17 1446 GCUUCUGGGCAGACUCUGA 2025UCAGAGUCUGCCCAGAAGC Rh [2002-2020] 17 1447 CCAGCCCUCUUCUGACACA 2026UGUGUCAGAAGAGGGCUGG [1846-1864] 17 1448 GCUCUAUCCCAACCUCUCA 2027UGAGAGGUUGGGAUAGAG Rh [1888-1906] 18 1449 GGACGUGGAGCGCACGGAA 2028UUCCGUGCGCUCCACGUCC Rh, D [799-817] 18 1450 CCAAGGCAGUGCUGAGCGA 2029UCGCUCAGCACUGCCUUGG Rh [507-525] 18 1451 GCAGAAGAAGGCUGUUGCA 2030UGCAACAGCCUUCUUCUGC Rt [1120-1138] 18 1452 GACAUUUUGUUGGAGCGUA 2031UACGCUCCAACAAAAUGUC [2183-2201] 18 1453 CGAGCACUCCAAGAUCAAA 2032UUUGAUCUUGGAGUGCUCG Rh, D [697-715] 18 1454 UCAUGAUGAUGCACCGGAA 2033UUCCGGUGCAUCAUCAUGA Rh [930-948] 18 1455 CCUGCUUCUCAGCGCCUUA 2034UAAGGCGCUGAGAAGCAGG [241-259] 18 1456 CCCAACCUCUCCCAACUAA 2035UUAGUUGGGAGAGGUUGG Rh [1895-1913] 18 1457 UGGGCAGACUCUGGUCAAA 2036UUUGACCAGAGUCUGCCCA Rh [2007-2025] 18 1458 CUCUGGUCAAGAAGCAUCA 2037UGAUGCUUCUUGACCAGAG Rh [2015-2033] 19 1459 GAGCCUCUCGAGCGCCUUA 2038UAAGGCGCUCGAGAGGCUC [1055-1073] 19 1460 AGAAGGCUGUUGCCAUCUA 2039UAGAUGGCAACAGCCUUCU Rt [1125-1143] 19 1461 CCCUGCUAGUCAACGCCAA 2040UUGGCGUUGACUAGCAGGG Rh [822-840] 19 1462 GCCUUCAGCUUGUACCAGA 2041UCUGGUACAAGCUGAAGGC [380-398] 19 1463 GCUGCUAACCAAAGAGCAA 2042UUGCUCUUUGGUUAGCAGC [1078-1096] 19 1464 CCCACAAGCUCUCCAGCCA 2043UGGCUGGAGAGCUUGUGGG Rh, D, P [1011-1029] 19 1465 GCUCCCUGCUAUUCAUUGA2044 UCAAUGAAUAGCAGGGAGC D [1422-1440] 19 1466 GUUCUUCAAAGAUAGGGAA 2045UUCCCUAUCUUUGAAGAAC [2083-2101] 19 1467 GUCAGCCAGCCCUCUUCUA 2046UAGAAGAGGGCUGGCUGAC Rh [1841-1859] 19 1468 GCGGGACACCCAAAGCGGA 2047UCCGCUUUGGGUGUCCCGC [1405-1423] 20 1469 AGCGCAGCGCGCUGCAGUA 2048UACUGCAGCGCGCUGCGCU Rh, Rt [726-744] 20 1470 CCGGAAACUCCACAUCCUA 2049UAGGAUGUGGAGUUUCCGG [1701-1719] 20 1471 CCAUUGACAAGAACAAGGA 2050UCCUUGUUCUUGUCAAUGG Rh, D [1215-1233] 20 1472 GGACAUCUACGGGCGCGAA 2051UUCGCGCCCGUAGAUGUCC D [1333-1351] 20 1473 GACACAUGGGUGCUAUUGA 2052UCAAUAGCACCCAUGUGUC Rh, Rt, M [1535-1553] 20 1474 CCUGGCACUGCGGAGAAGA2053 UCUUCUCCGCAGUGCCAGG [317-335] 20 1475 GGGCCUGACUGAGGCCAUA 2054UAUGGCCUCAGUCAGGCCC Rt [1201-1219] 20 1476 ACACUGGGAUGAGAAAUUA 2055UAAUUUCUCAUCCCAGUGU Rh [853-871] 20 1477 GGUCAGCCAGCCCUCUUCA 2056UGAAGAGGGCUGGCUGACC Rh [1840-1858] 20 1478 GUGAGGCGGAUUGAGAAGA 2057UCUUCUCAAUCCGCCUCAC [1969-1987] 21 1479 UCACCUGUGAGACCAAAUA 2058UAUUUGGUCUCACAGGUGA Rh [1811-1829] 21 1480 AGCUGCAAAUCGUGGAGAA 2059UUCUCCACGAUUUGCAGCU Rh  [984-1002] 21 1481 GGUGCACACAGGAUGGCAA 2060UUGCCAUCCUGUGUGCACC Rh [1495-1513] 21 1482 GGGUGUGGUGGAGGUGACA 2061UGUCACCUCCACCACACCC Rh, D [1153-1171] 21 1483 CCAGCCUUGGAUACUCCAA 2062UUGGAGUAUCCAAGGCUGG Rh [1575-1593] 21 1484 CCACAAGCUCUCCAGCCUA 2063UAGGCUGGAGAGCUUGUGG Rh, D, P [1012-1030] 21 1485 AAAGGCUCCUGAGACACAA2064 UUGUGUCUCAGGAGCCUUU [1523-1541] 21 1486 AGGAAAAGCUGCAAAUCGA 2065UCGAUUUGCAGCUUUUCCU Rh [978-996] 21 1487 CGCAGCAGCUCCUGGCACA 2066UGUGCCAGGAGCUGCUGCG [307-325] 21 1488 GGUGUCAUGAUGAUGCACA 2067UGUGCAUCAUCAUGACACC Rh [926-944] 22 1489 CCUCUUCUGACACUAAAAA 2068UUUUUAGUGUCAGAAGAG [1851-1869] 22 1490 AGCUAGAAUUCACUCCACA 2069UGUGGAGUGAAUUCUAGCU Rh [1649-1667] 22 1491 CGCUGGGCGGCAAGGCGAA 2070UUCGCCUUGCCGCCCAGCG [474-492] 22 1492 GGCCUGGCCUUCAGCUUGA 2071UCAAGCUGAAGGCCAGGCC [374-392] 22 1493 AGACACAUGGGUGCUAUUA 2072UAAUAGCACCCAUGUGUCU Rh, Rt, M [1534-1552] 22 1494 CGUGGGUGUCAUGAUGAUA2073 UAUCAUCAUGACACCCACG Rh [922-940] 22 1495 GUGGGUGUCAUGAUGAUGA 2074UCAUCAUCAUGACACCCAC Rh [923-941] 22 1496 GAGAAGGAGCUCCCAGGAA 2075UUCCUGGGAGCUCCUUCUC [1981-1999] 22 1497 GACUCUGGUCAAGAAGCAA 2076UUGCUUCUUGACCAGAGUC Rh [2013-2031] 22 1498 CACUAAAACACCUCAGCUA 2077UAGCUGAGGUGUUUUAGU [1861-1879] 23 1499 GGAGGCAUCCAAAGGCUCA 2078UGAGCCUUUGGAUGCCUCC [1513-1531] 23 1500 GACCCAGCUCAGUGAGCUA 2079UAGCUCACUGAGCUGGGUC [636-654] 23 1501 CCAUGACCUGCAGAAACAA 2080UUGUUUCUGCAGGUCAUGG Rh, Rt, M [1171-1189] 23 1502 AGAUGCAGAAGAAGGCUGA2081 UCAGCCUUCUUCUGCAUCU Rh, Rt, M [1116-1134] 23 1503CAGCAAGCAGCACUACAAA 2082 UUUGUAGUGCUGCUUGCUG Rh, D [676-694] 23 1504CAAGCUCUCCAGCCUCAUA 2083 UAUGAGGCUGGAGAGCUUG Rh, D, M, P [1015-1033] 231505 UGCAGAAGAAGGCUGUUGA 2084 UCAACAGCCUUCUUCUGCA Rt [1119-1137] 23 1506GGCGCGAGGAGCUGCGCAA 2085 UUGCGCAGCUCCUCGCGCC Rh, D, M [1344-1362] 231507 GGUACCAGCCUUGGAUACA 2086 UGUAUCCAAGGCUGGUACC Rh [1571-1589] 23 1508GCAGCCGACUGUACGGACA 2087 UGUCCGUACAGUCGGCUGC [621-639] 24 1509CAGCCUCAUCAUCCUCAUA 2088 UAUGAGGAUGAUGAGGCU Rh, D, Rt, [1024-1042] 241510 GCCACCGCCUUUGAGUUGA 2089 UCAACUCAAAGGCGGUGGC Rh [1289-1307] 24 1511AGAAGGACCUGUACCUGGA 2090 UCCAGGUACAGGUCCUUCU Rh, D [1257-1275] 24 1512GGUGAAGAAACCUGCAGCA 2091 UGCUGCAGGUUUCUUCACC Rh [289-307] 24 1513GUACCUUCUCACCUGUGAA 2092 UUCACAGGUGAGAAGGUAC Rh [1803-1821] 24 1514GGCCAAGGACCAGGCAGUA 2093 UACUGCCUGGUCCUUGGCC Rh [403-421] 24 1515GGCGGCAAGGCGACCACGA 2094 UCGUGGUCGCCUUGCCGCC [479-497] 24 1516AGCACUCCAAGAUCAACUA 2095 UAGUUGAUCUUGGAGUGCU Rh, D [699-717] 24 1517AUAUUUAUAGCCAGGUACA 2096 UGUACCUGGCUAUAAAUAU Rh [1789-1807] 24 1518GGCAGCCGACUGUACGGAA 2097 UUCCGUACAGUCGGCUGCC [620-638] 25 1519GUCACGCAUGUCAGGCAAA 2098 UUUGCCUGACAUGCGUGAC Rh, D [1240-1258] 25 1520GACAGGCCUCUACAACUAA 2099 UUAGUUGUAGAGGCCUGUC Rh, Rb, Rt, [946-964] 251521 GAUGCAGAAGAAGGCUGUA 2100 UACAGCCUUCUUCUGCAUC Rh, Rt, M [1117-1135]25 1522 ACCCAUGACCUGCAGAAAA 2101 UUUUCUGCAGGUCAUGGGU Rh, Rt, M[1169-1187] 25 1523 GGCUUCAUGGUGACUCGGA 2102 UCCGAGUCACCAUGAAGCC Rh[896-914] 25 1524 UGCCUCAAUCAGUAUUCAA 2103 UUGAAUACUGAUUGAGGCA[1772-1790] 25 1525 GUUCUUCAAGCCACACUGA 2104 UCAGUGUGGCUUGAAGAAC Rh, Rb,D [841-859] 25 1526 ACUCCAAGAUCAACUUCCA 2105 UGGAAGUUGAUCUUGGAG Rh, D,Rt, [702-720] 25 1527 GCUGUUCUACGCCGACCAA 2106 UUGGUCGGCGUAGAACAGC Rh[1369-1387] 25 1528 UAGUCAACGCCAUGUUCUA 2107 UAGAACAUGGCGUUGACUA Rh[828-846] 26 1529 CCGUGUGCCUGAGCGGACA 2108 UGUCCGCUCAGGCACACGG Rh[1625-1643] 26 1530 AGGCCUCUACAACUACUAA 2109 UUAGUAGUUGUAGAGGCCU Rh, Rb,D, [949-967] 26 1531 GCUUCAUGGUGACUCGGUA 2110 UACCGAGUCACCAUGAAGC Rh[897-915] 26 1532 GGUCAAGAAGCAUCGUGUA 2111 UACACGAUGCUUCUUGACC Rh[2019-2037] 26 1533 CUGCGAGCACUCCAAGAUA 2112 UAUCUUGGAGUGCUCGCAG Rh, D[694-712] 26 1534 GUCCUAUACCGUGGGUGUA 2113 UACACCCACGGUAUAGGAC Rh[913-931] 26 1535 GGCCUGACUGAGGCCAUUA 2114 UAAUGGCCUCAGUCAGGCC Rh[1202-1220] 26 1536 CACUCCAAGAUCAACUUCA 2115 UGAAGUUGAUCUUGGAGU Rh, D,Rt, [701-719] 26 1537 GCGUCGCAGGCCAAGGCAA 2116 UUGCCUUGGCCUGCGACGC[497-515] 26 1538 AAGGGUGACAAGAUGCGAA 2117 UUCGCAUCUUGUCACCCUU Rh, D[1457-1475] 27 1539 CAAGCUGUUCUACGCCGAA 2118 UUCGGCGUAGAACAGCUUG Rh[1366-1384] 27 1540 CCUGCUAGUCAACGCCAUA 2119 UAUGGCGUUGACUAGCAGG Rh[823-841] 27 1541 CCAAGGGUGUGGUGGAGGA 2120 UCCUCCACCACACCCUUGG Rh, D[1149-1167] 27 1542 CACACAGGAUGGCAGGAGA 2121 UCUCCUGCCAUCCUGUGUG Rh[1499-1517] 27 1543 UCCUGAGACACAUGGGUGA 2122 UCACCCAUGUGUCUCAGGA D, Rt,M [1529-1547] 27 1544 CUACAACUACUACGACGAA 2123 UUCGUCGUAGUAGUUGUAG Rb[955-973] 27 1545 GACAAGAUGCGAGACGAGA 2124 UCUCGUCUCGCAUCUUGUC Rh, Rt[1463-1481] 27 1546 CCUGGAAGCUGGGCAGCCA 2125 UGGCUGCCCAGCUUCCAGG[609-627] 27 1547 CUUCAAGCCACACUGGGAA 2126 UUCCCAGUGUGGCUUGAAG Rh, Rb, D[844-862] 27 1548 GCGAGACGAGUUAUAGGGA 2127 UCCCUAUAACUCGUCUCGC Rh[1471-1489] 28 1549 GAAGCUGGGCAGCCGACUA 2128 UAGUCGGCUGCCCAGCUUC[613-631] 28 1550 GUGCCUGAGCGGACCUUCA 7129 UGAAGGUCCGCUCAGGCAC Rh[1629-1647] 28 1551 GGUGACCCAUGACCUGCAA 2130 UUGCAGGUCAUGGGUCACC Rh, Rt,M [1165-1183] 28 1552 AUGAGCCUUUGUUGCUAUA 2131 UAUAGCAACAAAGGCUCAU Rh[2114-2132] 28 1553 CAACUACUACGACGACGAA 2132 UUCGUCGUCGUAGUAGUUG Rb[958-976] 28 1554 GCUGCGCUCACUCAGCAAA 2133 UUUGCUGAGUGAGCGCAGC Rh[571-589] 28 1555 GAGAACAUCCUGGUGUCAA 2134 UUGACACCAGGAUGUUCUC [422-440]28 1556 CCCAAGCUGUUCUACGCCA 2135 UGGCGUAGAACAGCUUGGG Rh [1364-1382] 281557 CAGCUCUAUCCCAACCUCA 2136 UGAGGUUGGGAUAGAGCU [1886-1904] 28 1558UGAGCUUCGCUGAUGACUA 2137 UAGUCAUCAGCGAAGCUCA Rh [648-666] 29 1559CCCAAGGCGGCCACGCUUA 2138 UAAGCGUGGCCGCCUUGGG Rh [341-359] 29 1560CUAUACCGUGGGUGUCAUA 2139 UAUGACACCCACGGUAUAG Rh [916-934] 29 1561CAUUGACAAGAACAAGGCA 2140 UGCCUUGUUCUUGUCAAUG Rh, D [1216-1234] 29 1562GGACCCAGCUCAGUGAGCA 2141 UGCUCACUGAGCUGGGUCC [635-653] 29 1563GACGACGAGAAGGAAAAGA 2142 UCUUUUCCUUCUCGUCGUC Rh [968-986] 29 1564GCGGCAAGGCGACCACGGA 2143 UCCGUGGUCGCCUUGCCGC [480-498] 29 1565GGGACACCCAAAGCGGCUA 2144 UAGCCGCUUUGGGUGUCCC [1407-1425] 29 1566GGGAGGUGAGGUACCAGCA 2145 UGCUGGUACCUCACCUCCC [1562-1580] 29 1567GCAGCACUACAACUGCGAA 2146 UUCGCAGUUGUAGUGCUGC Rh, D [682-700] 29 1568GCGCAACGUGACCUGGAAA 2147 UUUCCAGGUCACGUUGCGC M [598-616] 30 1569GGGCUGGGCCUGACUGAGA 2148 UCUCAGUCAGGCCCAGCCC [1196-1214] 30 1570CCUGAGCGGACCUUCCCAA 2149 UUGGGAAGGUCCGCUCAGG Rh [1632-1650] 30 1571GCAGCUGAAGAUCUGGAUA 2150 UAUCCAGAUCUUCAGCUGC Rh, D [1093-1111] 30 1572AGUGGAGAACAUCCUGGUA 2151 UACCAGGAUGUUCUCCACU Rh [418-436] 30 1573GCAAGCAGCACUACAACUA 2152 UAGUUGUAGUGCUGCUUGC Rh, D [678-696] 30 1574AGCUCAGUGAGCUUCGCUA 2153 UAGCGAAGCUCACUGAGCU [641-659] 30 1575CCGACUUGUCACGCAUGUA 2154 UACAUGCGUGACAAGUCGG Rh [1233-1251] 30 1576CCGAGGUCACCAAGGACGA 2155 UCGUCCUUGGUGACCUCGG Rh, D [786-804] 30 1577GGAGCCUCUCGAGCGCCUA 2156 UAGGCGCUCGAGAGGCUCC [1054-1072] 30 1578GGCCGCGCAGACCACCGAA 2157 UUCGGUGGUCUGCGCGGCC [757-775] 31 1579GGAAACUCCACAUCCUGUA 2158 UACAGGAUGUGGAGUUUCC Rh [1703-1721] 31 1580CAAAGCGGCUCCCUGCUAA 2159 UUAGCAGGGAGCCGCUUUG [1415-1433] 31 1581GCUCCUGAGACACAUGGGA 2160 UCCCAUGUGUCUCAGGAGC D [1527-1545] 31 1582CCUGGGCCAUAGUCAUUCA 2161 UGAAUGACUAUGGCCCAGG [1725-1743] 31 1583CGUGGAGCCUCUCGAGCGA 2162 UCGCUCGAGAGGCUCCACG [1051-1069] 31 1584CCUCCUGCUUCUCAGCGCA 2163 UGCGCUGAGAAGCAGGAGG [238-256] 31 1585AGUCCCAGAUCAAGCCUGA 2164 UCAGGCUUGAUCUGGGACU Rh [1756-1774] 31 1586UACCGUGGGUGUCAUGAUA 2165 UAUCAUGACACCCACGGUA Rh [919-937] 31 1587GCCAGCCCUCUUCUGACAA 2166 UUGUCAGAAGAGGGCUGGC [1845-1863] 31 1588CCGAGGUGAAGAAACCUGA 2167 UCAGGUUUCUUCACCUCGG Rh, Rt [285-303] 32 1589UCCUGGCACUGCGGAGAAA 2168 UUUCUCCGCAGUGCCAGGA [316-334] 32 1590CCCGGAAACUCCACAUCCA 2169 UGGAUGUGGAGUUUCCGGG [1700-1718] 32 1591ACUCUGGUCAAGAAGCAUA 2170 UAUGCUUCUUGACCAGAGU Rh [2014-2032] 32 1592CCCAGAUACCAUGAUGCUA 2171 UAGCAUCAUGGUAUCUGGG Rh [1679-1697] 32 1593CCUGAGACACAUGGGUGCA 2172 UGCACCCAUGUGUCUCAGG D, Rt, M [1530-1548] 321594 GCACUACAACUGCGAGCAA 2173 UUGCUCGCAGUUGUAGUGC Rh, D [685-703] 321595 CCACAAGAUGGUGGACAAA 2174 UUUGUCCACCAUCUUGUGG Rh, Rb, M, [874-892]32 1596 GGACACAGAUGGCAACCCA 2175 UGGGUUGCCAUCUGUGUCC [1306-1324] 32 1597GAAAAGCUGCUAACCAAAA 2176 UUUUGGUUAGCAGCUUUUC [1073-1091] 32 1598ACUACAACUGCGAGCACUA 2177 UAGUGCUCGCAGUUGUAGU Rh, D [687-705] 33 1599GCACUCCAAGAUCAACUUA 2178 UAAGUUGAUCUUGGAGUGC Rh, D [700-718] 33 1600GCCUUGAAAAGCUGCUAAA 2179 UUUAGCAGCUUUUCAAGGC [1068-1086] 33 1601GUGACUCGGUCCUAUACCA 2180 UGGUAUAGGACCGAGUCAC Rh [905-923] 33 1602GUGGUGGAGGUGACCCAUA 2181 UAUGGGUCACCUCCACCAC Rh, Rb, Rt, [1157-1175] 331603 AUGCGAGACGAGUUAUAGA 2182 UCUAUAACUCGUCUCGCAU Rh [1469-1487] 33 1604ACCUUCCCAGCUAGAAUUA 2183 UAAUUCUAGCUGGGAAGGU Rh [1641-1659] 33 1605CCCAGCUAGAAUUCACUCA 2184 UGAGUGAAUUCUAGCUGGG Rh [1646-1664] 33 1606GGUCACCAAGGACGUGGAA 2185 UUCCACGUCCUUGGUGACC Rh, D [790-808] 33 1607GGCCUCAGGGUGCACACAA 2186 UUGUGUGCACCCUGAGGCC [1487-1505] 33 1608UGAGGUACCAGCCUUGGAA 2187 UUCCAAGGCUGGUACCUCA Rh [1568-1586] 34 1609CAUGGUGACUCGGUCCUAA 2188 UUAGGACCGAGUCACCAUG Rh [901-919] 34 1610GGUGAGGUACCAGCCUUGA 2189 UCAAGGCUGGUACCUCACC Rh [1566-1584] 34 1611GCCGAGGUGAAGAAACCUA 2190 UAGGUUUCUUCACCUCGGC Rh, Rt [284-302] 34 1612GUACGGACCCAGCUCAGUA 2191 UACUGAGCUGGGUCCGUAC [631-649] 34 1613CAAGAAGGACCUGUACCUA 2192 UAGGUACAGGUCCUUCUUG Rh, D, M [1255-1273] 341614 GAGCACUCCAAGAUCAACA 2193 UGUUGAUCUUGGAGUGCUC Rh, D [698-716] 341615 CAUGUUCUUCAAGCCACAA 2194 UUGUGGCUUGAAGAACAUG Rh, Rb, D [838-856] 341616 CCCUCCUGCUUCUCAGCGA 2195 UCGCUGAGAAGCAGGAGGG [237-255] 34 1617AUGUCAGGCAAGAAGGACA 2196 UGUCCUUCUUGCCUGACAU Rh, D [1247-1265] 34 1618CAAGAUCAACUUCCGCGAA 2197 UUCGCGGAAGUUGAUCUUG D [706-724] 35 1619GCGUGUUCCACGCCACCGA 2198 UCGGUGGCGUGGAACACGC [1278-1296] 35 1620CGGACCCAGCUCAGUGAGA 2199 UCUCACUGAGCUGGGUCCG [634-652] 35 1621CCUUCAGCUUGUACCAGGA 2200 UCCUGGUACAAGCUGAAGG [381-399] 35 1622GCUCUCCAGCCUCAUCAUA 2201 UAUGAUGAGGCUGGAGAGC Rh, D, Rt, [1018-1036] 351623 CCCUGGCCCACAAGCUCUA 2202 UAGAGCUUGUGGGCCAGGG Rh, D, P [1005-1023]35 1624 GCCCGAGGUCACCAAGGAA 2203 UUCCUUGGUGACCUCGGGC Rh, D [784-802] 351625 GUGGAGAACAUCCUGGUGA 2204 UCACCAGGAUGUUCUCCAC Rh [419-437] 35 1626GCUCACUCAGCAACUCCAA 2205 UUGGAGUUGCUGAGUGAGC Rh [576-594] 35 1627ACGCCAUGUUCUUCAAGCA 2206 UGCUUGAAGAACAUGGCGU Rh, Rb, P [834-852] 35 1628ACACAUGGGUGCUAUUGGA 2907 UCCAAUAGCACCCAUGUGU Rh [1536-1554] 36 1629CCAGCUCAGUGAGCUUCGA 2208 UCGAAGCUCACUGAGCUGG [639-657] 36 1630CCCAGCUCAGUGAGCUUCA 2209 UGAAGCUCACUGAGCUGGG [638-656] 36 1631GGGCGGCAAGGCGACCACA 2210 UGUGGUCGCCUUGCCGCCC [478-496] 36 1632CAGGGUGCACACAGGAUGA 2211 UCAUCCUGUGUGCACCCUG [1492-1510] 36 1633AGGUGAAGAAACCUGCAGA 2212 UCUGCAGGUUUCUUCACCU Rh [288-306] 36 1634CCUCUCCCAACUAUAAAAA 2213 UUUUUAUAGUUGGGAGAG Rh [1900-1918] 36 1635GACUGUACGGACCCAGCUA 2214 UAGCUGGGUCCGUACAGUC [627-645] 36 1636GAAGGAGCUCCCAGGAGGA 2215 UCCUCCUGGGAGCUCCUUC [1983-2001] 36 1637ACGCAUGUCAGGCAAGAAA 2216 UUUCUUGCCUGACAUGCGU Rh, D [1243-1261] 36 1638GACUCGGUCCUAUACCGUA 2217 UACGGUAUAGGACCGAGUC Rh [907-925] 37 1639CACUACAACUGCGAGCACA 2218 UGUGCUCGCAGUUGUAGUG Rh, D [686-704] 37 1640AGCUCCUGGCACUGCGGAA 2219 UUCCGCAGUGCCAGGAGCU [313-331] 37 1641CUAAGGGUGACAAGAUGCA 2220 UGCAUCUUGUCACCCUUAG Rh [1455-1473] 37 1642UGUGAGACCAAAUUGAGCA 2221 UGCUCAAUUUGGUCUCACA Rh [1816-1834] 37 1643GCCGACUUGUCACGCAUGA 2222 UCAUGCGUGACAAGUCGGC Rh [1232-1250] 37 1644CAGGAUGGCAGGAGGCAUA 2223 UAUGCCUCCUGCCAUCCUG [1503-1521] 37 1645ACAAGAACAAGGCCGACUA 2224 UAGUCGGCCUUGUUCUUGU Rh [1221-1239] 37 1646UGCGCUCCCUCCUGCUUCA 2225 UGAAGCAGGAGGGAGCGCA [231-249] 37 1647GGCGAGCUGCUGCGCUCAA 2226 UUGAGCGCAGCAGCUCGCC Rh [563-581] 37 1648GAUGCACCGGACAGGCCUA 2227 UAGGCCUGUCCGGUGCAUC Rh, Rb, Rt, [937-955] 381649 CGUGUCGCUGGGCGGCAAA 2228 UUUGCCGCCCAGCGACACG [469-487] 38 1650AUCCCAACCUCUCCCAACA 2229 UGUUGGGAGAGGUUGGGA Rh [1893-1911] 38 1651UGUUCUACGCCGACCACCA 2230 UGGUGGUCGGCGUAGAACA Rh [1371-1389] 38 1652CGGCCUGGCCUUCAGCUUA 2231 UAAGCUGAAGGCCAGGCCG [373-391] 38 1653GUCGCAGGCCAAGGCAGUA 2232 UACUGCCUUGGCCUGCGAC [499-517] 38 1654AGUCAUUCUGCCUGCCCUA 2233 UAGGGCAGGCAGAAUGACU [1735-1753] 38 1655CCCAGAAUGACCUGGCCGA 2234 UCGGCCAGGUCAUUCUGGG [1949-1967] 38 1656ACAAGAUGGUGGACAACCA 2235 UGGUUGUCCACCAUCUUGU Rh, Rb, M, [876-894] 381657 GCUAGUCAACGCCAUGUUA 2236 UAACAUGGCGUUGACUAGC Rh [826-844] 38 1658ACGCCACCGCCUUUGAGUA 2237 UACUCAAAGGCGGUGGCGU Rh [1287-1305] 39 1659GCCGCGCAGACCACCGACA 2238 UGUCGGUGGUCUGCGCGGC [758-776] 39 1660GCUAUUCAUUGGGCGCCUA 2239 UAGGCGCCCAAUGAAUAGC D [1429-1447] 39 1661CUCAGUGAGCUUCGCUGAA 2240 UUCAGCGAAGCUCACUGAG [643-661] 39 1662GGAGGUGAGGUACCAGCCA 2241 UGGCUGGUACCUCACCUCC [1563-1581] 39 1663GCCAAGGCAGUGCUGAGCA 2242 UGCUCAGCACUGCCUUGGC Rh [506-524] 39 1664CUCUCCAGCCUCAUCAUCA 2243 UGAUGAUGAGGCUGGAGA Rh, D, Rt, [1019-1037] 391665 GAAUGACCUGGCCGCAGUA 2244 UACUGCGGCCAGGUCAUUC [1953-1971] 39 1666UGGUGACUCGGUCCUAUAA 2245 UUAUAGGACCGAGUCACCA Rh [903-921] 39 1667CAGGUACCUUCUCACCUGA 2246 UCAGGUGAGAAGGUACCUG Rh [1800-1818] 39 1668GUUCCACGCCACCGCCUUA 2247 UAAGGCGGUGGCGUGGAAC D [1282-1300] 40 1669CCGACUGUACGGACCCAGA 2248 UCUGGGUCCGUACAGUCGG [625-643] 40 1670GCAGACCACCGACGGCAAA 2249 UUUGCCGUCGGUGGUCUGC D, Rt [763-781] 40 1671AAGAUGCGAGACGAGUUAA 2250 UUAACUCGUCUCGCAUCUU Rh [1466-1484] 40 1672CAAAGAGCAGCUGAAGAUA 2251 UAUCUUCAGCUGCUCUUUG Rh [1087-1105] 40 1673ACGACGAGAAGGAAAAGCA 2252 UGCUUUUCCUUCUCGUCGU Rh [969-987] 40 1674CACUCCACUUGGACAUGGA 2253 UCCAUGUCCAAGUGGAGUG Rh [1659-1677] 40 1675AGUCCAUCAACGAGUGGGA 2254 UCCCACUCGUUGAUGGACU Rh, Rt, M [741-759] 40 1676GCGCCGGCCUGGCCUUCAA 2255 UUGAAGGCCAGGCCGGCGC Rh [369-387] 40 1677GGAAAAGCUGCAAAUCGUA 2256 UACGAUUUGCAGCUUUUCC Rh [979-997] 40 1678ACAUUUUGUUGGAGCGUGA 2257 UCACGCUCCAACAAAAUGU [2184-2202] 41 1679ACCGUGGCUUCAUGGUGAA 2258 UUCACCAUGAAGCCACGGU Rh, Rt, M [891-909] 41 1680CCCUUCAUCUUCCUAGUGA 2259 UCACUAGGAAGAUGAAGGG [1388-1406] 41 1681GAAAUUCCACCACAAGAUA 2260 UAUCUUGUGGUGGAAUUUC Rh [865-883] 41 1682CUAUAAAACUAGGUGCUGA 2261 UCAGCACCUAGUUUUAUAG Rh [1910-1928] 41 1683GGAGGUGCACGCCGGCCUA 2262 UAGGCCGGCGUGCACCUCC [544-562] 41 1684GCAGGCCAAGGCAGUGCUA 2263 UAGCACUGCCUUGGCCUGC [502-520] 41 1685UGAGACCAAAUUGAGCUAA 2264 UUAGCUCAAUUUGGUCUCA Rh [1818-1836] 41 1686GCCAUAGUCAUUCUGCCUA 2265 UAGGCAGAAUGACUAUGGC [1730-1748] 41 1687AGCUGAAGAUCUGGAUGGA 2266 UCCAUCCAGAUCUUCAGCU Rh, D [1095-1113] 41 1688CCAUCUCCUUGCCCAAGGA 2267 UCCUUGGGCAAGGAGAUGG Rh [1137-1155] 42 1689CCCAGAUCAAGCCUGCCUA 2268 UAGGCAGGCUUGAUCUGGG Rh [1759-1777] 42 1690GCUGUUGCCAUCUCCUUGA 2269 UCAAGGAGAUGGCAACAGC [1130-1148] 42 1691CGAGGUCACCAAGGACGUA 2270 UACGUCCUUGGUGACCUCG Rh, D [787-805] 42 1692CAACUAUAAAACUAGGUGA 2271 UCACCUAGUUUUAUAGUUG Rh [1907-1925] 42 1693GAAGGCUGUUGCCAUCUCA 2272 UGAGAUGGCAACAGCCUUC Rt [1126-1144] 42 1694UGCGGAGAAGUUGAGCCCA 2273 UGGGCUCAACUUCUCCGCA [325-343] 42 1695CUCCUUGCCCAAGGGUGUA 2274 UACACCCUUGGGCAAGGAG Rh [1141-1159] 42 1696GCCCUGAAAGUCCCAGAUA 2275 UAUCUGGGACUUUCAGGGC [1748-1766] 42 1697CAAGGGUGUGGUGGAGGUA 2276 UACCUCCACCACACCCUUG Rh, D [1150-1168] 42 1698AAGAGCAGCUGAAGAUCUA 2277 UAGAUCUUCAGCUGCUCUU Rh [1089-1107] 43 1699GAAGAUGCAGAAGAAGGCA 2278 UGCCUUCUUCUGCAUCUUC Rh, Rb, Rt [1114-1132] 431700 CGGAAACUCCACAUCCUGA 2279 UCAGGAUGUGGAGUUUCCG [1702-1720] 43 1701AGUCAACGCCAUGUUCUUA 2280 UAAGAACAUGGCGUUGACU Rh [829-847] 43 1702CGAGCGCCUUGAAAAGCUA 2281 UAGCUUUUCAAGGCGCUCG [1063-1081] 43 1703AUACCGUGGGUGUCAUGAA 2282 UUCAUGACACCCACGGUAU Rh [918-936] 43 1704GACCUGGGCCAUAGUCAUA 2283 UAUGACUAUGGCCCAGGUC [1723-1741] 43 1705CAUGUCAGGCAAGAAGGAA 2284 UUCCUUCUUGCCUGACAUG Rh, D [1246-1264] 43 1706UGCGAGACGAGUUAUAGGA 2285 UCCUAUAACUCGUCUCGCA Rh [1470-1488] 43 1707CGCAACGUGACCUGGAAGA 2286 UCUUCCAGGUCACGUUGCG [599-617] 43 1708AGCAAGCAGCACUACAACA 2287 UGUUGUAGUGCUGCUUGCU Rh, D [677-695] 44 1709GCUGCUGCGCUCACUCAGA 2288 UCUGAGUGAGCGCAGCAGC Rh [568-586] 44 1710UGAUGAUGCACCGGACAGA 2289 UCUGUCCGGUGCAUCAUCA Rh [933-951] 44 1711UUGUUGCUAUCAAUCCAAA 2290 UUUGGAUUGAUAGCAACAA Rh [2122-2140] 44 1712CCUUGAAAAGCUGCUAACA 2291 UGUUAGCAGCUUUUCAAGG [1069-1087] 44 1713CCCUUUGACCAGGACAUCA 2292 UGAUGUCCUGGUCAAAGGG Rh, Rt [1322-1340] 44 1714GAGGUGAAGAAACCUGCAA 2293 UUGCAGGUUUCUUCACCUC Rh [287-305] 44 1715CCCAAGGGUGUGGUGGAGA 2294 UCUCCACCACACCCUUGGG Rh, D [1148-1166] 44 1716CCCUGCUAUUCAUUGGGCA 2295 UGCCCAAUGAAUAGCAGGG D [1425-1443] 44 1717CUGAAAGUCCCAGAUCAAA 2296 UUUGAUCUGGGACUUUCAG [1751-1769] 44 1718GCUGCAAAUCGUGGAGAUA 2297 UAUCUCCACGAUUUGCAGC Rh  [985-1003] 45 1719CAAGCCUGCCUCAAUCAGA 2298 UCUGAUUGAGGCAGGCUUG Rh [1766-1784] 45 1720CGAGCAGCUGCGCGACGAA 2299 UUCGUCGCGCAGCUGCUCG [526-544] 45 1721AGGCCGACUUGUCACGCAA 2300 UUGCGUGACAAGUCGGCCU Rh [1230-1248] 45 1722GCAGCAGCUCCUGGCACUA 2301 UAGUGCCAGGAGCUGCUGC [308-326] 45 1723GGCCAUAGUCAUUCUGCCA 2302 UGGCAGAAUGACUAUGGCC [1729-1747] 45 1724CCCGUGUGCCUGAGCGGAA 2303 UUCCGCUCAGGCACACGGG Rh [1624-1642] 45 1725CAGCUGAAGAUCUGGAUGA 2304 UCAUCCAGAUCUUCAGCUG Rh, D [1094-1112] 45 1726CAAGCCACACUGGGAUGAA 2305 UUCAUCCCAGUGUGGCUUG Rh, Rb [847-865] 45 1727GAAUUCACUCCACUUGGAA 2306 UUCCAAGUGGAGUGAAUUC Rh [1654-1672] 45 1728CGGCGCCCUGCUAGUCAAA 2307 UUUGACUAGCAGGGCGCCG Rh [817-835] 46 1729UGGAAGCUGGGCAGCCGAA 2308 UUCGGCUGCCCAGCUUCCA [611-629] 46 1730GGCAAGGCGACCACGGCGA 2309 UCGCCGUGGUCGCCUUGCC Rh [482-500] 46 1731CACUGCGGAGAAGUUGAGA 2310 UCUCAACUUCUCCGCAGUG [322-340] 46 1732GGCAGGAGGCAUCCAAAGA 2311 UCUUUGGAUGCCUCCUGCC [1509-1527] 46 1733GGUGACUCGGUCCUAUACA 2312 UGUAUAGGACCGAGUCACC Rh [904-922] 46 1734UUUAUAGCCAGGUACCUUA 2313 UAAGGUACCUGGCUAUAAA Rh [1792-1810] 46 1735GGCCAUGGCCAAGGACCAA 2314 UUGGUCCUUGGCCAUGGCC Rh, D [397-415] 46 1736CAAAGAUAGGGAGGGAAGA 2315 UCUUCCCUCCCUAUCUUUG [2089-2107] 46 1737UCUUCUGACACUAAAACAA 2316 UUGUUUUAGUGUCAGAAG [1853-1871] 46 1738CUUCUGACACUAAAACACA 2317 UGUGUUUUAGUGUCAGAA [1854-1872] 47 1739UCACGUGGAGCCUCUCGAA 2318 UUCGAGAGGCUCCACGUGA [1048-1066] 47 1740CAGUCCAUCAACGAGUGGA 2319 UCCACUCGUUGAUGGACUG Rh, Rt, M [740-758] 47 1741AGACCAAAUUGAGCUAGGA 2320 UCCUAGCUCAAUUUGGUCU [1820-1838] 47 1742GGGUUCCCGUGUGCCUGAA 2321 UUCAGGCACACGGGAACCC Rh [1619-1637] 47 1743UUGCUAUCAAUCCAAGAAA 2322 UUUCUUGGAUUGAUAGCAA Rh [2125-2143] 47 1744CAACCGUGGCUUCAUGGUA 2323 UACCAUGAAGCCACGGUUG Rh, Rt, M [889-907] 47 1745CUGUACGGACCCAGCUCAA 2324 UUGAGCUGGGUCCGUACAG [629-647] 47 1746CAGCAGCAAGCAGCACUAA 2325 UUAGUGCUGCUUGCUGCUG Rh, D [673-691] 47 1747CCUGCAGCCGCAGCAGCUA 2326 UAGCUGCUGCGGCUGCAGG [299-317] 47 1748GACACUAAAACACCUCAGA 2327 UCUGAGGUGUUUUAGUGUC [1859-1877] 48 1749CAACUGCGAGCACUCCAAA 2328 UUUGGAGUGCUCGCAGUUG Rh, D [691-709] 48 1750ACUGCGGAGAAGUUGAGCA 2329 UGCUCAACUUCUCCGCAGU [323-341] 48 1751GCGCCCUGCUAGUCAACGA 2330 UCGUUGACUAGCAGGGCGC Rh [819-837] 48 1752GGAAGCUGGGCAGCCGACA 2331 UGUCGGCUGCCCAGCUUCC [612-630] 48 1753AGGCUCCUGAGACACAUGA 2332 UCAUGUGUCUCAGGAGCCU D [1525-1543] 48 1754CGACAAGCGCAGCGCGCUA 2333 UAGCGCGCUGCGCUUGUCG [721-739] 48 1755UCAGUGAGCUUCGCUGAUA 2334 UAUCAGCGAAGCUCACUGA [644-662] 48 1756UUGAGAAGGAGCUCCCAGA 2335 UCUGGGAGCUCCUUCUCAA [1979-1997] 48 1757ACUGCGAGCACUCCAAGAA 2336 UUCUUGGAGUGCUCGCAGU Rh, D [693-711] 48 1758CAUCCUGGUGUCACCCGUA 2337 UACGGGUGACACCAGGAUG [427-445] 49 1759GUGCGCAGCAGCAAGCAGA 2338 UCUGCUUGCUGCUGCGCAC Rh, D [668-686] 49 1760CACGCCACCGCCUUUGAGA 2339 UCUCAAAGGCGGUGGCGUG Rh [1286-1304] 49 1761UCUCGAGCGCCUUGAAAAA 2340 UUUUUCAAGGCGCUCGAGA [1060-1078] 49 1762GCUUCGCUGAUGACUUCGA 2341 UCGAAGUCAUCAGCGAAGC Rh [651-669] 49 1763UCUCCUUGCCCAAGGGUGA 2342 UCACCCUUGGGCAAGGAGA Rh [1140-1158] 49 1764GCAGUCCAUCAACGAGUGA 2343 UCACUCGUUGAUGGACUGC Rh, Rt, M [739-757] 49 1765AGAUGGUGGACAACCGUGA 2344 UCACGGUUGUCCACCAUCU Rh, M [879-897] 49 1766CGGCUCCCUGCUAUUCAUA 2345 UAUGAAUAGCAGGGAGCCG [1420-1438] 49 1767AUACCAUGAUGCUGAGCCA 2346 UGGCUCAGCAUCAUGGUAU [1684-1702] 49 1768AGCCAGGUACCUUCUCACA 2347 UGUGAGAAGGUACCUGGCU Rh [1797-1815] 50 1769GAGCCCGGAAACUCCACAA 2348 UUGUGGAGUUUCCGGGCUC [1697-1715] 50 1770GCAGCUCCUGGCACUGCGA 2349 UCGCAGUGCCAGGAGCUGC [311-329] 50 1771CCCGAGGUCACCAAGGACA 2350 UGUCCUUGGUGACCUCGGG Rh, D [785-803] 50 1772CCUGACUGAGGCCAUUGAA 2351 UUCAAUGGCCUCAGUCAGG Rh [1204-1222] 50 1773UGCUGAGCCCGGAAACUCA 2352 UGAGUUUCCGGGCUCAGCA [1693-1711] 50 1774GCCAUCUCCUUGCCCAAGA 2353 UCUUGGGCAAGGAGAUGGC Rh [1136-1154] 50 1775CAAGCAGCACUACAACUGA 2354 UCAGUUGUAGUGCUGCUUG Rh, D [679-697] 50 1776CAAGGCAGUGCUGAGCGCA 2355 UGCGCUCAGCACUGCCUUG Rh [508-526] 50 1777CAAUGACAUUUUGUUGGAA 2356 UUCCAACAAAAUGUCAUUG [2179-2197] 50 1778AGUGAGCUUCGCUGAUGAA 2357 UUCAUCAGCGAAGCUCACU [646-664] 51 1779AUGAUGAUGCACCGGACAA 2358 UUGUCCGGUGCAUCAUCAU Rh [932-950] 51 1780GAAACACCUGGCUGGGCUA 2359 UAGCCCAGCCAGGUGUUUC D [1183-1201] 51 1781CCUGCUAUUCAUUGGGCGA 2360 UCGCCCAAUGAAUAGCAGG D [1426-1444] 51 1782CGCCACCGCCUUUGAGUUA 2361 UAACUCAAAGGCGGUGGCG Rh [1288-1306] 51 1783GCUUCUCAGCGCCUUCUGA 2362 UCAGAAGGCGCUGAGAAGC [244-262] 51 1784UGAUGCUGAGCCCGGAAAA 2363 UUUUCCGGGCUCAGCAUCA [1690-1708] 51 1785UGACCUGGCCGCAGUGAGA 2364 UCUCACUGCGGCCAGGUCA [1956-1974] 51 1786UGCAGAAACACCUGGCUGA 2365 UCAGCCAGGUGUUUCUGCA [1179-1197] 51 1787GCAGUGCUGAGCGCCGAGA 2366 UCUCGGCGCUCAGCACUGC [512-530] 51 1788CGGCGCGCAACGUGACCUA 2367 UAGGUCACGUUGCGCGCCG [594-612] 52 1789AGUGCUGAGCGCCGAGCAA 2368 UUGCUCGGCGCUCAGCACU [514-532] 52 1790ACAGGCCUCUACAACUACA 2369 UGUAGUUGUAGAGGCCUGU Rh, Rb, D, [947-965] 521791 GCAGCUGCGCGACGAGGAA 2370 UUCCUCGUCGCGCAGCUGC Rh, D [529-547] 521792 AUUGAGAAGGAGCUCCCAA 2371 UUGGGAGCUCCUUCUCAAU [1978-1996] 52 1793CGCGCAGACCACCGACGGA 2372 UCCGUCGGUGGUCUGCGCG [760-778] 52 1794CCUGUACCUGGCCAGCGUA 2373 UACGCUGGCCAGGUACAGG Rh [1264-1282] 52 1795CUGAGCGGACCUUCCCAGA 2374 UCUGGGAAGGUCCGCUCAG Rh [1633-1651] 52 1796GGCCUUCAGCUUGUACCAA 2375 UUGGUACAAGCUGAAGGCC [379-397] 52 1797CACCCAAAGCGGCUCCCUA 2376 UAGGGAGCCGCUUUGGGUG [1411-1429] 52 1798GCCAAGGACCAGGCAGUGA 2377 UCACUGCCUGGUCCUUGGC Rh [404-422] 53 1799CUCAGGGUGCACACAGGAA 2378 UUCCUGUGUGCACCCUGAG [1490-1508] 53 1800CGAGCUGCUGCGCUCACUA 2379 UAGUGAGCGCAGCAGCUCG Rh [565-583] 53 1801GGCUGGGCCUGACUGAGGA 2380 UCCUCAGUCAGGCCCAGCC [1197-1215] 53 1802CCGCAGCAGCUCCUGGCAA 2381 UUGCCAGGAGCUGCUGCGG [306-324] 53 1803UGUGGGACCUGGGCCAUAA 2382 UUAUGGCCCAGGUCCCACA [1718-1736] 53 1804AAGAUGCAGAAGAAGGCUA 2383 UAGCCUUCUUCUGCAUCUU Rh, Rt, M [1115-1133] 531805 CCACGGCGCGCAACGUGAA 2384 UUCACGUUGCGCGCCGUGG Rh [591-609] 53 1806ACCUUCUCACCUGUGAGAA 2385 UUCUCACAGGUGAGAAGGU Rh [1805-1823] 53 1807UGAAGAAACCUGCAGCCGA 2386 UCGGCUGCAGGUUUCUUCA [291-309] 53 1808CAGCACUACAACUGCGAGA 2387 UCUCGCAGUUGUAGUGCUG Rh, D [683-701] 54 1809GCGACAAGCGCAGCGCGCA 2388 UGCGCGCUGCGCUUGUCGC [720-738] 54 1810UAGAAUUCACUCCACUUGA 2389 UCAAGUGGAGUGAAUUCUA Rh [1652-1670] 54 1811GUGGAAAAACAGACCGGGA 2390 UCCCGGUCUGUUUUUCCAC [1603-1621] 54 1812ACGUGGAGCCUCUCGAGCA 2391 UGCUCGAGAGGCUCCACGU [1050-1068] 54 1813GGCGCGCAACGUGACCUGA 2392 UCAGGUCACGUUGCGCGCC [595-613] 54 1814UGGACAACCGUGGCUUCAA 2393 UUGAAGCCACGGUUGUCCA Rh, M [885-903] 54 1815CUAGUCAACGCCAUGUUCA 2394 UGAACAUGGCGUUGACUAG Rh [827-845] 54 1816AGAAUGACCUGGCCGCAGA 2395 UCUGCGGCCAGGUCAUUCU [1952-1970] 54 1817AGCUGCUGCGCUCACUCAA 2396 UUGAGUGAGCGCAGCAGCU Rh [567-585] 54 1818CUCUAUCCCAACCUCUCCA 2397 UGGAGAGGUUGGGAUAGA Rh [1889-1907] 55 1819GCGAGCUGCUGCGCUCACA 2398 UGUGAGCGCAGCAGCUCGC Rh [564-582] 55 1820CGCAGCAGCAAGCAGCACA 2399 UGUGCUGCUUGCUGCUGCG Rh, D [671-689] 55 1821GGCUGGGCUGGGCCUGACA 2400 UGUCAGGCCCAGCCCAGCC [1192-1210] 55 1822UCUCCAGCCUCAUCAUCCA 2401 UGGAUGAUGAGGCUGGAG Rh, D, Rt, [1020-1038] 551823 CAACGCCAUGUUCUUCAAA 2402 UUUGAAGAACAUGGCGUUG Rh, Rb, P [832-850] 551824 UGGCACUGCGGAGAAGUUA 2403 UAACUUCUCCGCAGUGCCA [319-337] 55 1825UUUGAGUUGGACACAGAUA 2404 UAUCUGUGUCCAACUCAAA [1298-1316] 55 1826UGGGCGAGCUGCUGCGCUA 2405 UAGCGCAGCAGCUCGCCCA Rh [561-579] 55 1827CUGCUAACCAAAGAGCAGA 2406 UCUGCUCUUUGGUUAGCAG [1079-1097] 55 1828AACGUGACCUGGAAGCUGA 2407 UCAGCUUCCAGGUCACGUU [602-620] 56 1829AUGACAUUUUGUUGGAGCA 2408 UGCUCCAACAAAAUGUCAU [2181-2199] 56 1830CAGGAGGCAUCCAAAGGCA 2409 UGCCUUUGGAUGCCUCCUG [1511-1529] 56 1831AUCUCCUUGCCCAAGGGUA 2410 UACCCUUGGGCAAGGAGAU Rh [1139-1157] 56 1832UGGGAUGAGAAAUUCCACA 2411 UGUGGAAUUUCUCAUCCCA Rh [857-875] 56 1833AAAGCUGCUAACCAAAGAA 2412 UUCUUUGGUUAGCAGCUUU [1075-1093] 56 1834AGGAGGCAUCCAAAGGCUA 2413 UAGCCUUUGGAUGCCUCCU [1512-1530] 56 1835CACCGCCUUUGAGUUGGAA 2414 UUCCAACUCAAAGGCGGUG Rh [1291-1309] 56 1836CCAACUAUAAAACUAGGUA 2415 UACCUAGUUUUAUAGUUGG Rh [1906-1924] 56 1837CAAGAAGCAUCGUGUCUGA 2416 UCAGACACGAUGCUUCUUG Rh [2022-2040] 56 1838AGCAGCUGAAGAUCUGGAA 2417 UUCCAGAUCUUCAGCUGCU Rh, D [1092-1110] 57 1839GCGCUCCCUCCUGCUUCUA 2418 UAGAAGCAGGAGGGAGCGC [232-250] 57 1840UGCUAGUCAACGCCAUGUA 2419 UACAUGGCGUUGACUAGCA Rh [825-843] 57 1841CGCCGAGCAGCUGCGCGAA 2420 UUCGCGCAGCUGCUCGGCG [523-541] 57 1842CCGCGCAGACCACCGACGA 2421 UCGUCGGUGGUCUGCGCGG [759-777] 57 1843UAGCCAGGUACCUUCUCAA 2422 UUGAGAAGGUACCUGGCUA Rh [1796-1814] 57 1844UGCUUCUCAGCGCCUUCUA 2423 UAGAAGGCGCUGAGAAGCA [243-261] 57 1845CUCCCUCCUGCUUCUCAGA 2424 UCUGAGAAGCAGGAGGGAG [235-253] 57 1846CGCAGGCCAAGGCAGUGCA 2425 UGCACUGCCUUGGCCUGCG [501-519] 57 1847GCAAGGCGACCACGGCGUA 2426 UACGCCGUGGUCGCCUUGC Rh [483-501] 57 1848GCAGCCGCAGCAGCUCCUA 2427 UAGGAGCUGCUGCGGCUGC [302-320]

TABLE E SERPINH1 Cross-Species 18 + 1-mer siRNAs No. SEQ ID NO SensesiRNA SEQ AntiSense siRNA Other human- 1 2428 UCACCAAGGACGUGGAGCA 2576UGCUCCACGUCCUUGGU Rh, D [792-810] 2 2429 CAGCGCGCUGCAGUCCAUA 2577UAUGGACUGCAGCGCGC Rh, Rt [730-748] 3 2430 CAUCUACGGGCGCGAGGAA 2578UUCCUCGCGCCCGUAGA D, M [1336-1354] 4 2431 CUCCAGCCUCAUCAUCCUA 2579UAGGAUGAUGAGGCUGG Rh, D, Rt, M [1021-1039] 5 2432 GACAUCUACGGGCGCGAGA2580 UCUCGCGCCCGUAGAUG D, M [1334-1352] 6 2433 CGUGCGCAGCAGCAAGCAA 2581UUGCUUGCUGCUGCGCA Rh, D, M [667-685] 7 2434 GUCACCAAGGACGUGGAGA 2582UCUCCACGUCCUUGGUG Rh, D [791-809] 8 2435 CCGCGACAAGCGCAGCGCA 2583UGCGCUGCGCUUGUCGC D [718-736] 9 2436 GCGCAGCGCGCUGCAGUCA 2584UGACUGCAGCGCGCUGC Rh, Rt [727-745] 10 2437 GGCCCACAAGCUCUCCAGA 2585UCUGGAGAGCUUGUGGG Rh, D, P [1009-1027] 11 2438 CAAGGACGUGGAGCGCACA 2586UGUGCGCUCCACGUCCU Rh, D [796-814] 12 2439 AGCCUCAUCAUCCUCAUGA 2587UCAUGAGGAUGAUGAGG Rh, D, Rt, M [1025-1043] 13 2440 GGUGUGGUGGAGGUGACCA2588 UGGUCACCUCCACCACA Rh, D [1154-1172] 14 2441 GCAAGCUGCCCGAGGUCAA2589 UUGACCUCGGGCAGCUU Rh, D [777-795] 15 2442 GUGGAGGUGACCCAUGACA 2590UGUCAUGGGUCACCUCC Rh, Rt, M [1160-1178] 16 2443 CACAAGAUGGUGGACAACA 2591UGUUGUCCACCAUCUUG Rh, Rb, M, P [875-893] 17 2444 GCGAGGAGCUGCGCAGCCA2592 UGGCUGCGCAGCUCCUC D, M [1347-1365] 18 2445 UACUACGACGACGAGAAGA 2593UCUUCUCGUCGUCGUAG Rb [962-980] 19 2446 GAGGUGACCCAUGACCUGA 2594UCAGGUCAUGGGUCACC Rh, Rt, M [1163-1181] 20 2447 ACUUCCGCGACAAGCGCAA 2595UUGCGCUUGUCGCGGAA D [714-732] 21 2448 GCCCACAAGCUCUCCAGCA 2596UGCUGGAGAGCUUGUGG Rh, D, P [1010-1028] 22 2449 GCGCAGCAGCAAGCAGCAA 2597UUGCUGCUUGCUGCUGC Rh, D [670-688] 23 2450 CGAGGAGCUGCGCAGCCCA 2598UGGGCUGCGCAGCUCCU D, M [1348-1366] 24 2451 AACGCCAUGUUCUUCAAGA 2599UCUUGAAGAACAUGGCG Rh, Rb, P [833-851] 25 2452 GUCAGGCAAGAAGGACCUA 2600UAGGUCCUUCUUGCCUG Rh, D [1249-1267] 26 2453 GCCUGGGCGAGCUGCUGCA 2601UGCAGCAGCUCGCCCAG Rh, D [558-576] 27 2454 GAUGAUGCACCGGACAGGA 2602UCCUGUCCGGUGCAUCA Rh, Rb, Rt, M [934-952] 28 2455 GGACCUGUACCUGGCCAGA2603 UCUGGCCAGGUACAGGU Rh, D [1261-1279] 29 2456 GCGACGAGGAGGUGCACGA2604 UCGUGCACCUCCUCGUC D [537-555] 30 2457 UGUGGUGGAGGUGACCCAA 2605UUGGGUCACCUCCACCA Rh, D [1156-1174] 31 2458 UUCAAGCCACACUGGGAUA 2606UAUCCCAGUGUGGCUUG Rh, Rb [845-863] 32 2459 CAAGAUGGUGGACAACCGA 2607UCGGUUGUCCACCAUCU Rh, Rb, M, P [877-895] 33 2460 UCAACUUCCGCGACAAGCA2608 UGCUUGUCGCGGAAGUU D [711-729] 34 2461 AUUCAUUGGGCGCCUGGUA 2609UACCAGGCGCCCAAUGA D [1432-1450] 35 2462 CUCCAAGAUCAACUUCCGA 2610UCGGAAGUUGAUCUUGG Rh, D, Rt, M [703-721] 36 2463 CAGGCCAUGGCCAAGGACA2611 UGUCCUUGGCCAUGGCC Rh, D [395-413] 37 2464 GUACCAGGCCAUGGCCAAA 2612UUUGGCCAUGGCCUGGU Rh, D [391-409] 38 2465 UGUCAGGCAAGAAGGACCA 2613UGGUCCUUCUUGCCUGA Rh, D [1248-1266] 39 2466 CUUCGUGCGCAGCAGCAAA 2614UUUGCUGCUGCGCACGA Rh, D, M [664-682] 40 2467 CAACUUCCGCGACAAGCGA 2615UCGCUUGUCGCGGAAGU D [712-730] 41 2468 CCACCACAAGAUGGUGGAA 2616UUCCACCAUCUUGUGGU Rh, Rb, D, P [871-889] 42 2469 GCGCGACGAGGAGGUGCAA2617 UUGCACCUCCUCGUCGC Rh, D [535-553] 43 2470 CUACAACUGCGAGCACUCA 2618UGAGUGCUCGCAGUUGU Rh, D [688-706] 44 2471 UGGAGGUGACCCAUGACCA 2619UGGUCAUGGGUCACCUC Rh, Rt, M [1161-1179] 45 2472 GAGGUCACCAAGGACGUGA 2620UCACGUCCUUGGUGACC Rh, D [788-806] 46 2473 AAGAAGGACCUGUACCUGA 2621UCAGGUACAGGUCCUUC Rh, D [1256-1274] 47 2474 GACAACCGUGGCUUCAUGA 2622UCAUGAAGCCACGGUUG Rh, Rt, M [887-905] 48 2475 ACCAGGACAUCUACGGGCA 2623UGCCCGUAGAUGUCCUG D, Rt [1329-1347] 49 2476 GCUGCCCGAGGUCACCAAA 2624UUUGGUGACCUCGGGCA Rh, D [781-799] 50 2477 AUGCAGAAGAAGGCUGUUA 2625UAACAGCCUUCUUCUGC Rt [1118-1136] 51 2478 GGCCUGGGCGAGCUGCUGA 2626UCAGCAGCUCGCCCAGG Rh, D [557-575] 52 2479 GAUGGUGGACAACCGUGGA 2627UCCACGGUUGUCCACCA Rh, M [880-898] 53 2480 CUCCCUGCUAUUCAUUGGA 2628UCCAAUGAAUAGCAGGG D [1423-1441] 54 2481 GAAGGACCUGUACCUGGCA 2629UGCCAGGUACAGGUCCU Rh, D [1258-1276] 55 2482 CCACCGACGGCAAGCUGCA 2630UGCAGCUUGCCGUCGGU D, Rt [768-786] 56 2483 UGCUAUUCAUUGGGCGCCA 2631UGGCGCCCAAUGAAUAG D [1428-1446] 57 2484 AUGUUCUUCAAGCCACACA 2632UGUGUGGCUUGAAGAAC Rh, Rb, D [839-857] 58 2485 CCAGGACAUCUACGGGCGA 2633UCGCCCGUAGAUGUCCU D, Rt [1330-1348] 59 2486 GCGCGAGGAGCUGCGCAGA 2634UCUGCGCAGCUCCUCGC Rh, D, M [1345-1363] 60 2487 GAGCAGCUGCGCGACGAGA 2635UCUCGUCGCGCAGCUGC Rh, D [527-545] 61 2488 CUAUUCAUUGGGCGCCUGA 2636UCAGGCGCCCAAUGAAU D [1430-1448] 62 2489 ACAAGCUCUCCAGCCUCAA 2637UUGAGGCUGGAGAGCUU Rh, D, M, P [1014-1032] 63 2490 GCUGAAGAUCUGGAUGGGA2638 UCCCAUCCAGAUCUUCA Rh, D [1096-1114] 64 2491 GACCAGGACAUCUACGGGA2639 UCCCGUAGAUGUCCUGG D, Rt [1328-1346] 65 2492 CAAGCGCAGCGCGCUGCAA2640 UUGCAGCGCGCUGCGCU Rh, Rt [724-742] 66 2493 CCAUGGCCAAGGACCAGGA 2641UCCUGGUCCUUGGCCAU Rh, D [399-417] 67 2494 CACCAAGGACGUGGAGCGA 2642UCGCUCCACGUCCUUGG Rh, D [793-811] 68 2495 CCGUGGCUUCAUGGUGACA 2643UGUCACCAUGAAGCCAC Rh, Rt, M [892-910] 69 2496 UGACCAGGACAUCUACGGA 2644UCCGUAGAUGUCCUGGU Rt [1327-1345] 70 2497 AGACCACCGACGGCAAGCA 2645UGCUUGCCGUCGGUGGU D, Rt [765-783] 71 2498 GACAAGCGCAGCGCGCUGA 2646UCAGCGCGCUGCGCUUG Rh, Rt [722-740] 72 2499 AGAAACACCUGGCUGGGCA 2647UGCCCAGCCAGGUGUUU D [1182-1200] 73 2500 AAGAUGGUGGACAACCGUA 2648UACGGUUGUCCACCAUC Rh, M [878-896] 74 2501 CAGACCACCGACGGCAAGA 2649UCUUGCCGUCGGUGGUC D, Rt [764-782] 75 2502 AGGACCUGUACCUGGCCAA 2650UUGGCCAGGUACAGGUC Rh, D [1260-1278] 76 2503 CUGCUAUUCAUUGGGCGCA 2651UGCGCCCAAUGAAUAGC D [1427-1445] 77 2504 GUCCAUCAACGAGUGGGCA 2652UGCCCACUCGUUGAUGG Rh, Rt, M [742-760] 78 2505 CCAGGCCAUGGCCAAGGAA 2653UUCCUUGGCCAUGGCCU Rh, D [394-412] 79 2506 AAGCAGCACUACAACUGCA 2654UGCAGUUGUAGUGCUGC Rh, D [680-698] 80 2507 UGUUCCACGCCACCGCCUA 2655UAGGCGGUGGCGUGGAA D [1281-1299] 81 2508 UACAACUACUACGACGACA 2656UGUCGUCGUAGUAGUUG Rb [956-974] 82 2509 CCUCAUCAUCCUCAUGCCA 2657UGGCAUGAGGAUGAUGA Rh, D, Rt, M [1027-1045] 83 2510 UGGUGGACAACCGUGGCUA2658 UAGCCACGGUUGUCCAC Rh, M [882-900] 84 2511 GACCACCGACGGCAAGCUA 2659UAGCUUGCCGUCGGUGG D, Rt [766-784] 85 2512 AGCUGCGCGACGAGGAGGA 2660UCCUCCUCGUCGCGCAG Rh, D [531-549] 86 2513 CGGCAAGCUGCCCGAGGUA 2661UACCUCGGGCAGCUUGC Rh, D [775-793] 87 2514 UGGCCCACAAGCUCUCCAA 2662UUGGAGAGCUUGUGGGC Rh, D, P [1008-1026] 88 2515 CAGCUGCGCGACGAGGAGA 2663UCUCCUCGUCGCGCAGC Rh, D [530-548] 89 2516 CUUCCGCGACAAGCGCAGA 2664UCUGCGCUUGUCGCGGA D [715-733] 90 2517 UGGGCCUGACUGAGGCCAA 2665UUGGCCUCAGUCAGGCC Rt [1200-1218] 91 2518 GCUGCGCGACGAGGAGGUA 2666UACCUCCUCGUCGCGCA Rh, D [532-550] 92 2519 CAGGACAUCUACGGGCGCA 2667UGCGCCCGUAGAUGUCC D [1331-1349] 93 2520 GCCAUGCCGAAGGACCAGA 2668UCUGGUCCUUGGCCAUG Rh, D [398-416] 94 2521 UCCAAGAUCAACUUCCGCA 2669UGCGGAAGUUGAUCUUG D [704-722] 95 2592 ACCACCGACGGCAAGCUGA 2670UCAGCUUGCCGUCGGUG D, Rt [767-785] 96 2523 AUCUACGGGCGCGAGGAGA 2671UCUCCUCGCGCCCGUAG D, M [1337-1355] 97 2524 CUGCCCGAGGUCACCAAGA 2672UCUUGGUGACCUCGGGC Rh, D [782-800] 98 2525 AUCAACUUCCGCGACAAGA 2673UCUUGUCGCGGAAGUUG D [710-728] 99 2526 UCAUUGGGCGCCUGGUCCA 2674UGGACCAGGCGCCCAAU Rh, D [1434-1452] 100 2527 CAUUGGGCGCCUGGUCCGA 2675UCGGACCAGGCGCCCAA Rh, D [1435-1453] 101 2528 GUGUUCCACGCCACCGCCA 2676UGGCGGUGGCGUGGAAC D [1280-1298] 102 2529 AUGAUGCACCGGACAGGCA 2677UGCCUGUCCGGUGCAUC Rh, Rb, Rt, M [935-953] 103 2530 CGACGAGGAGGUGCACGCA2678 UGCGUGCACCUCCUCGU D [538-556] 104 2531 CAGAAACACCUGGCUGGGA 2679UCCCAGCCAGGUGUUUC D [1181-1199] 105 2532 UGAUGCACCGGACAGGCCA 2680UGGCCUGUCCGGUGCAU Rh, Rb, Rt, M [936-954] 106 2533 AAGGCUGUUGCCAUCUCCA2681 UGGAGAUGGCAACAGCC D, Rt [1127-1145] 107 2534 AUGACUUCGUGCGCAGCAA2682 UUGCUGCGCACGAAGUC Rh, Rt, M [660-678] 108 2535 UCAGGCAAGAAGGACCUGA2683 UCAGGUCCUUCUUGCCU Rh, D [1250-1268] 109 2536 CUCAUCAUCCUCAUGCCCA2684 UGGGCAUGAGGAUGAUG Rh, Rt, M [1028-1046] 110 2537CGCGACGAGGAGGUGCACA 2685 UGUGCACCUCCUCGUCG Rh, D [536-554] 111 2538ACAACCGUGGCUUCAUGGA 2686 UCCAUGAAGCCACGGUU Rh, Rt, M [888-906] 112 2539UUGACCAGGACAUCCACGA 2687 UCGUAGAUGUCCUGGUC Rt [1326-1344] 113 2540CAAGCUGCCCGAGGUCACA 2688 UGUGACCUCGGGCAGCU Rh, D [778-796] 114 2541UCCCUGCUAUUCAUUGGGA 2689 UCCCAAUGAAUAGCAGG D [1424-1442] 115 2542UAUUCAUUGGGCGCCUGGA 2690 UCCAGGCGCCCAAUGAA D [1431-1449] 116 2543CUGCGCGACGAGGAGGUGA 2691 UCACCUCCUCGUCGCGC Rh, D [533-551] 117 2544CUACGGGCGCGAGGAGCUA 2692 UAGCUCCUCGCGCCCGU D, M [1339-1357] 118 2545CGCGAGGAGCUGCGCAGCA 2693 UGCUGCGCAGCUCCUCG D, M [1346-1364] 119 2546ACACCUGGCUGGGCUGGGA 2694 UCCCAGCCCAGCCAGGU D [1186-1204] 120 2547UCUACGGGCGCGAGGAGCA 2695 UGCUCCUCGCGCCCGUA D, M [1338-1356] 121 2548UUCUUCAAGCCACACUGGA 2696 UCCAGUGUGGCUUGAAG Rh, Rb, D [842-860] 122 2549CCUGGGCGAGCUGCUGCGA 2697 UCGCAGCAGCUCGCCCA Rh, D [559-577] 123 2550AAGAAGGCUGUUGCCAUCA 2698 UGAUGGCAACAGCCUUC Rt [1124-1142] 124 2551CGACGGCAAGCUGCCCGAA 2699 UUCGGGCAGCUUGCCGU D [772-790] 125 2552GACGGCAAGCUGCCCGAGA 2700 UCUCGGGCAGCUUGCCG Rh, D [773-791] 126 2553UUCAUUGGGCGCCUGGUCA 2701 UGACCAGGCGCCCAAUG Rh, D [1433-1451] 127 2554AAGCGCAGCGCGCUGCAGA 2702 UCUGCAGCGCGCUGCGC Rh, Rt [725-743] 128 2555CCUGGCCCACAAGCUCUCA 2703 UGAGAGCUUGUGGGCCA Rh, D, P [1006-1024] 129 2556ACGGCAAGCUGCCCGAGGA 2704 UCCUCGGGCAGCUUGCC Rh, D [774-792] 130 2557UUUGACCAGGACAUCUACA 2705 UGUAGAUGUCCUGGUCA Rt [1325-1343] 131 2558UGACUUCGUGCGCAGCAGA 2706 UCUGCUGCGCACGAAGU Rh, Rt, M [661-679] 132 2559AAGGACGUGGAGCGCACGA 2707 UCGUGCGCUCCACGUCC Rh, D [797-815] 133 2560UCCAUCAACGAGUGGGCCA 2708 UGGCCCACUCGUUGAUG Rt, M [743-761] 134 2561CACCGACGCCAACCUGCCA 2709 UGGCAGCUUGCCGUCGG D, Rt [769-787] 135 2562ACGGGCGCGAGGAGCUGCA 2710 UGCAGCUCCUCGCGCCC D, M [1341-1359] 136 2563UCCGCGACAAGCGCAGCGA 2711 UCGCUGCGCUUGUCGCG D [717-735] 137 2564UUGGGCGCCUGGUCCGGCA 2712 UGCCGGACCAGGCGCCC Rh, D [1437-1455] 138 2565AUGGUGGACAACCGUGGCA 2713 UGCCACGGUUGUCCACC Rh, M [881-899] 139 2566AUUGGGCGCCUGGUCCGGA 2714 UCCGGACCAGGCGCCCA Rh, D [1436-1454] 140 2567UACGGGCGCGAGGAGCUGA 2715 UCAGCUCCUCGCGCCCG D, M [1340-1358] 141 2568AUGCACCGGACAGGCCUCA 2716 UGAGGCCUGUCCGGUGC Rh, Rb, Rt, P [938-956] 1422569 UUCCACCACAAGAUGGUGA 2717 UCACCAUCUUGUGGUGG Rh, Rb, D, P [869-887]143 2570 UUCCGCGACAAGCGCAGCA 2718 UGCUGCGCUUGUCGCGG D [716-734] 144 2571UACCAGGCCAUGGCCAAGA 2719 UCUUGGCCAUGGCCUGG Rh, D [392-410] 145 2572AAACACCUGGCUGGGCUGA 2720 UCAGCCCAGCCAGGUGU D [1184-1202] 146 2573ACCGACGGCAAGCUGCCCA 2721 UGGGCAGCUUGCCGUCG D [770-788] 147 2574AACACCUGGCUGGGCUGGA 2722 UCCAGCCCAGCCAGGUG D [1185-1203] 148 2575UUCGUGCGCAGCAGCAAGA 2723 UCUUGCUGCUGCGCACG Rh, D, M [665-683]

Example 10 Animal Models

Model Systems of Fibrotic Conditions

Testing the active siRNAs of the invention may be done in predictiveanimal models. Rat diabetic and aging models of kidney fibrosis includeZucker diabetic fatty (ZDF) rats, aged falfa (obese Zucker) rats, agedSprague-Dawley (SD) rats, and Goto Kakizaki (GK) rats; GK rats are aninbred strain derived from Wistar rats, selected for spontaneousdevelopment of NIDDM (diabetes type II). Induced models of kidneyfibrosis include the permanent unilateral ureteral obstruction (UUO)model which is a model of acute interstitial fibrosis occurring inhealthy non-diabetic animals; renal fibrosis develops within daysfollowing the obstruction. Another induced model of kidney fibrosis is5/6 nephrectomy.

Two models of liver fibrosis in rats are the Bile Duct Ligation (BDL)with sham operation as controls, and CCl₄ poisoning, with olive oil fedanimals as controls, as described in the following references:Lotersztajn S, et al Hepatic Fibrosis: Molecular Mechanisms and DrugTargets. Annu Rev Pharmacol Toxicol. 2004 Oct. 7; Uchio K, et al.,Down-regulation of connective tissue growth factor and type I collagenmRNA expression by connective tissue growth factor antisenseoligonucleotide during experimental liver fibrosis. Wound Repair Regen.2004 January-February; 12(1):60-6; Xu X Q, et al., Molecularclassification of liver cirrhosis in a rat model by proteomics andbioinformatics Proteomics. 2004 October; 4(10):3235-45.

Models for ocular scarring are well known in the art e.g. Sherwood M Bet al., J. Glaucoma. 2004 October; 13(5):407-12. A new model of glaucomafiltering surgery in the rat; Miller M H et al., Ophthalmic Surg. 1989May; 20(5):350-7. Wound healing in an animal model of glaucomafistulizing surgery in the Rb; vanBockxmeer F M et al., Retina. 1985Fall-Winter; 5(4): 239-52. Models for assessing scar tissue inhibitors;Wiedemann P et al., J Pharmacol Methods. 1984 August; 12(1): 69-78.Proliferative vitreoretinopathy: the Rb cell injection model forscreening of antiproliferative drugs.

Models of cataract are described in the following publications: The roleof Src family kinases in cortical cataract formation. Zhou J, Menko AS.Invest Ophthalmol V is Sci. 2002 July; 43(7):2293-300; Bioavailabilityand anticataract effects of a topical ocular drug delivery systemcontaining disulfuram and hydroxypropyl-beta-cyclodextrin onselenite-treated rats. Wang S, et al.http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids-15370367 Curr Eye Res. 2004 July; 29(1):51-8; and Long-termorgan culture system to study the effects of UV-A irradiation on lenstransglutaminase. Weinreb O, Dovrat A.; Curr Eye Res. 2004 July;29(1):51-8.

The compounds of Table A-18 and Table A-19 are tested in these models offibrotic conditions, in which it is found that they are effective intreating liver fibrosis and other fibrotic conditions.

Model Systems of Glaucoma

Testing the active siRNA of the invention for treating or preventingglaucoma is preformed in rat animal model for optic nerve crushdescribed for example in: Maeda, K. et al., “A Novel Neuroprotectantagainst Retinal Ganglion Cell Damage in a Glaucoma Model and an OpticNerve Crush Model in the rat”, Investigative Ophthalmology and visualScience (IOVS), March 2004, 45(3)851. Specifically, for optic nervetransection the orbital optic nerve (ON) of anesthetized rats is exposedthrough a supraorbital approach, the meninges severed and all axons inthe ON transected by crushing with forceps for 10 seconds, 2 mm from thelamina cribrosa.

Nucleic acid molecules as disclosed herein are tested in this animalmodel and the results show that these siRNA compounds are useful intreating and/or preventing glaucoma.

Rat Optic Nerve Crush (ONC) Model: Intravitreal siRNA Delivery and EyeDrop Delivery

For optic nerve transsection the orbital optic nerve (ON) ofanesthetized rats is exposed through a supraorbital approach, themeninges severed and all axons in the ON transected by crushing withforceps for 10 seconds, 2 mm from the lamina cribrosa.

The siRNA compounds are delivered alone or in combination in 5 uL volume(10 ug/uL) as eye drops. Immediately after optic nerve crush (ONC), 20ug/10 ul test siRNA or 10 ul PBS is administered to one or both eyes ofadult Wistar rats and the levels of siRNA taken up into the dissectedand snap frozen whole retinae at 5 h and 1 d, and later at 2 d, 4 d, 7d, 14 d and 21 d post injection is determined. Similar experiments areperformed in order to test activity and efficacy of siRNA administeredvia eye drops.

Model Systems of Ischemia Reperfusion Injury Following LungTransplantation in Rats

Lung ischemia/reperfusion injury is achieved in a rat animal model asdescribed in Mizobuchi et al., The Journal of Heart and LungTransplantation, Vol 23 No. 7 (2004) and in Kazuhiro Yasufuku et al.,Am. J. Respir. Cell Mol Biol, Vol 25, pp 26-34 (2001).

Specifically, after inducing anesthesia with isofluorane, the trachea iscannulated with a 14-gauge Teflon catheter and the rat is mechanicallyventilated with rodent ventilator using 100% oxygen, at a rate of 70breaths per minute and 2 cm H2O of positive end-respiratory pressure.The left pulmonary artery, veins and main stem bronchus are occludedwith a Castaneda clamp. During the operation, the lung is kept moistwith saline and the incision is covered to minimize evaporative losses.The period of ischemia is 60 minutes long. At the end of the ischemicperiod the clamp is removed and the lung is allowed to ventilate andreperfuse for further 4 h, 24 h, and 5 d post induction of lungischemia. At the end of the experiment, the lungs are gently harvestedand either frozen for RNA extraction or fixed in glutaraldehyde cocktailfor subsequent histological analysis.

The Bleomycin Animal Model as a Model for Idiopathic Pulmonary Fibrosis(IPF).

Testing feasibility of lung and liver delivery of vitamin A-Coatsomeformulated siRNA administered by intravenous injection and intratrachealadministration of siRNA-vitamin A-Coatsome complex to a healthy mice andbleomycine-treated mice

Objective: To test two administration routes for feasibility of vitaminA-Coatsome formulated siRNA delivery to normal and fibrotic mouse lungs.The main hypothesis to be tested in the current study is whethersystemic administration of vitamin A-Coatsome formulated modified siRNAprovides efficient uptake and cell-specific distribution in the fibroticand normal mouse lungs. Intratracheal route of vitamin A-Coatsomeformulated modified siRNA will be tested in parallel. siRNA detectionand cell-specific distribution in the lungs and liver will be performedby in situ hybridization (ISH)

Background: The Bleomycin model of pulmonary fibrosis has been welldeveloped and characterized over the last three decades (Moeller, et al.Int J Biochem Cell Biol, 40:362-382, 2008; Chua et al., Am J Respir CellMol Biol 33:9-13, 2005). Histological hallmarks, such as intra-alveolarbuds, mural incorporation of collagen and obliteration of alveolar spaceare present in BLM-treated animals similar to IPF patients. Earlystudies demonstrated that C57/B1 mice were consistently prone toBLM-induced lung fibrosis, whereas Balb/C mice were inheritantlyresistant. Depending on the route of administration, different fibroticpattern develops. Intratracheal instillation of BLM results inbronchiocentric accentuated fibrosis, whereas intravenous orintraperitoneal administration induces subpleural scarring similar tohuman disease (Chua et al. ibid). A mouse model of usual interstitialpneumonia (UIP) is used. This model shows a heterogenous distribution offibroproliferation, distributed mainly subpleurally, forming similarlesions to those observed in the lungs of patients with idiopathicpulmonary fibrsosis (IPF) (Onuma, et al., Tohoku J Exp Med 194: 147-156,2001 and Yamaguchi and Ruoslahti, Nature 336: 244-246, 1988). UIP willbe induced by intraperitoneal injection of bleomycin every other day for7 days for a constant composition of subpleural fibroproliferation inthe mouse lung (Swiderski et al. Am J Pathol 152: 821-828, 1998 andShimizukawa et al., Am J Physiol Lung Cell Mol Physiol 284: L526-L532,2003).

As was previously demonstrated, vitamin A-loaded liposomes containingsiRNA interact with retinol-binding protein (RBP) and provide efficientdelivery to the hepatic stellate cells via RBP receptor (Sato et al. NatBiotechnol 26:431-442, 2008). This study is planned to test whethervitA-Coatsome-siRNA complex will be efficiently taken up by an RBPreceptor-expressing activated myofibroblasts in the lungs ofbleomycin-treated mice. In addition, local administration route(intratracheal instillation) will be tested.

General Study Design

Mice—C57 B1 maleStarting N (BLM I.P.)—40 (6 for the first pilot group, 34 for the study,taling in consideration anticipated 25% mortality)

Starting N (Total)—60

Test siRNA: SERPINHI compounds disclosed herein.

Groups: BLM dose, Termination, mg/kg siRNA post BW, in BLM dose, siRNAlast N (before 0.1 ml adm. BLM mg/kg adm siRNA siRNA siRNA No salineroute regime BW route regime adm administration 1 0.75 I.P. dd 0, 2, 4,6 4.5 I.V. 2 daily  2 h 4 2 0.75 I.P. dd 0, 2, 4, 6 4.5 I.V. 2 daily 24h 4 3 0.75 I.P. dd 0, 2, 4, 6 2.25 I.T. 2 daily  2 h 4 4 0.75 I.P. dd 0,2, 4, 6 2.25 I.T. 2 daily 24 h 4 5 intact n/a 4.5 I.V. 2 daily  2 h 4 6intact n/a 4.5 I.V 2 daily 24 h 4 7 intact n/a 2.25 I.T 2 daily  2 h 4 8intact n/a 2.25 I.T. 2 daily 24 h 4 9 0.75 I.P. dd 0, 2, 4, 6 n/a I.V. 2daily  2 h 3 vehicle 10 0.75 I.P. dd 0, 2, 4, 6 n/a I/T/vehicle 2 daily24 h 3 11 Intact n/a n/a intact n/a Any time 3

Bleomycin-induced pulmonary fibrosis. Pulmonary fibrosis of 12-wk-oldfemale C57BL/6 mice will be induced by intraperitoneal instillation ofbleomycin chlorate: 0.75 mg/kg body weight dissolved in 0.1 ml of salineevery other day for 7 days, on days 0, 2, 4, and 6.

Model follow up and monitoring. The mice will be weighed before the BLMtreatment, and twice weekly for the period of study duration.

Pilot evaluation of the establishment of fibrosis. The mice (N=30) aresubjected to BLM treatment in groups, to allow for a one week timeinterval between the first treated group (N=5) and the rest of theanimals. On day 14, two mice from the first group are sacrificed and thelungs harvested for the fast HE stain and quick histopathologicalevaluation of fibrosis. When lung fibrosis is confirmed, the remainingrats are sorted into the groups and treated with siRNA on Day 14 afterthe first BLM treatment. In case that no sufficient fibrosis develops inthe lungs by day 14, the remaining mice from the first treated group aresacrificed on day 21, followed by quick histopathology evaluation offibrosis. The rest of the animals are treated with test siRNA complexstarting from day 21 after the BLM treatment.

siRNA administration. On day 14 or day 21 after the first BLMadministration (TBD during the study, based on pilot evaluation ofestablishment of fibrosis), the animals are group sorted, according toBW. The animals from groups 1 and 2 are administered intravenously (tailvein injection) with siRNA/vitA/Coatsome complex, at an siRNAconcentration of 4.5 mg/kg BW. Intact animals of the same age (Groups 5and 6) are treated in the same manner. BLM treated animals (Group 9)will be used as vitA-coatsome vehicle control). In 24 hours, theinjection is repeated to all the animals, as above.

The BLM animals from the groups 3 and 4, and intact mice from groups 7and 8 are anesthetized with isoflutrane and subjected to intratrachealinstillation of 2.25 mg/kg BW siRNA formulated in vitA-loaded liposomes.Mice from the BLM group 10 are administered with vitA/Coatsome vehicleonly. The inttratracheal instillation is repeated after 24 hours.

Study termination. The animals from the groups 1, 3, 5, 7, 9 aresacrificed at 2 hours after the second siRNA complex injection orinstillation. The animals from the groups 2,4,6,8,10 are sacrificed at24 hours after the second siRNA complex injection or instillation.

Upon animals sacrifice, the mice are perfused transcardially with 10%neutral buffered formalin. The lungs are inflated with 0.8-1.0 ml of 10%NBF, and the trachea ligated. The lungs are excised and fixed for 24 hin 10% NBF. The liver is harvested from each animal and fixed in 10% NBFfor 24 h.

Sectioning and evaluation. Consequent sections are prepared from thelungs and livers. First consequent section are stained with hematoxylinand eosin for assessment of lung and liver morphology, second sectionare stained with Sirius Red (trichrome) to identify collagen The thirdconsequent sections are subjected to in situ hybridization (ISH) fordetection of siRNA.

The compounds as described herein are tested in this animal model andthe results show that these siRNA compounds are useful in treatingand/or preventing ischemia reperfusion injury following lungtransplantation.

TABLE 1 List of siRNA sequences Experimental Target Base sequence[corresponding sequence [corresponding siRNA region nucleotides of SEQID NO: 1] nucleotides of SEQ ID NO: 1] siHSP47-C human/rat sense5′ GGACAGGCCUCUACAACUAUU (SEQ ID 5′ GGACAGGCCUCUACAACUAdTdT (SEQ hsp47NO: 3) ID NO: 5) [945-963] [945-963] anti- 5′ UAGUUGUAGAGGCCUGUCCUU (SEQ5′ UAGUUGUAGAGGCCUGUCCdTdT (SEQ sense ID NO: 4) ID NO: 6) [945-963][945-963] siHSP47- human/rat sense 5′ GGACAGGCCUCUACAACUACUACGA5′ GGACAGGCCUCUACAACUACUACdGdA Cd hsp47 (SEQ ID NO: 7) (SEQ ID NO: 9)[945-969] [945-969] anti- 5′ 5′ sense UCGUAGUAGUUGUAGAGGCCUGUCCUUUCGUAGUAGUUGUAGAGGCCUGUCCUU (SEQ ID NO: 8) (SEQ ID NO: 10) [945-969][945-969] siHSP47-1 human/rat sense 5′ CAGGCCUCUACAACUACUAUU (SEQ ID5′ CAGGCCUCUACAACUACUAdTdT (SEQ hsp47 NO: 11) ID NO: 13) [948-966][948-966] anti- 5′ UAGUAGUUGUAGAGGCCUGUU (SEQ 5′ UAGUAGUUGUAGAGGCCUGdTdT(SEQ sense ID NO: 12) ID NO: 14) [948-966] [948-966] siHSP47- humansense 5′ CAGGCCUCUACAACUACUACGACGA 5′ CAGGCCUCUACAACUACUACGACdGdA 1dhsp47 (SEQ ID NO: 15) (SEQ ID NO: 17) [948-972] [948-972] anti- 5′ 5′sense UCGUCGUAGUAGUUGUAGAGGCCUGUU UCGUCGUAGUAGUUGUAGAGGCCUGUU (SEQ IDNO: 16) (SEQ ID NO: 18) [948-972] [948-972] siHsp47-2 human sense5′ GAGCACUCCAAGAUCAACUUU (SEQ ID 5′ GAGCACUCCAAGAUCAACUdTdT (SEQ hsp47NO: 19) ID NO: 21) [698-717] [698-717] anti- 5′ AGUUGAUCUUGGAGUGCUCUU(SEQ 5′ AGUUGAUCUUGGAGUGCUCdTdT (SEQ sense ID NO: 20) ID NO: 22)[698-716] [698-716] siHsp47-2d human sense 5′ GAGCACUCCAAGAUCAACUUCCGCG5′ GAGCACUCCAAGAUCAACUUCCGdCdG hsp47 (SEQ ID NO: 23) (SEQ ID NO: 25)[698-722] [698-722] anti- 5′ 5′ sense CGCGGAAGUUGAUCUUGGAGUGCUCUUCGCGGAAGUUGAUCUUGGAGUGCUCUU (SEQ ID NO: 24) (SEQ ID NO: 26) [698-722][698-722] siHsp47-2d rat Gp46 sense 5′ GAACACUCCAAGAUCAACUUCCGAG5′ GAACACUCCAAGAUCAACUUCCGdAdG rat (SEQ ID NO: 27) (SEQ ID NO: 29)[587-611] [587-611] anti- 5′ 5′ sense CUCGGAAGUUGAUCUUGGAGUGUUCUUCUCGGAAGUUGAUCUUGGAGUGUUCUU (SEQ ID NO: 28) (SEQ ID NO: 30) [587-611][587-611] siHsp47-3 human sense 5′ CUGAGGCCAUUGACAAGAAUU (SEQ5′ CUGAGGCCAUUGACAAGAAdTdT (SEQ hsp47 ID NO: 31) ID NO: 33) cDNA[1209-1227] [1209-1227] anti- 5′ UUCUUGUCAAUGGCCUCAGUU (SEQ5′ UUCUUGUCAAUGGCCUCAGdTdT (SEQ sense ID NO: 32) ID NO: 34) [1209-1227][1209-1227] siHsp47-3d human sense 5′ CUGAGGCCAUUGACAAGAACAAGGC 5′ hsp47(SEQ ID NO: 35) CUGAGGCCAUUGACAAGAACAAGdGdC [1209-1233] (SEQ ID NO: 37)[1209-1233] anti- 5′ 5′ sense GCCUUGUUCUUGUCAAUGGCCUCAGUUGCCUUGUUCUUGUCAAUGGCCUCAGUU (SEQ ID NO: 36) (SEQ ID NO: 38) [1209-1233][1209-1233] siHsp47-4 human sense 5′ CUACGACGACGAGAAGGAAUU (SEQ5′ CUACGACGACGAGAAGGAAdTdT (SEQ hsp47 ID NO: 39) ID NO: 41) [964-982][964-982] anti- 5′ UUCCUUCUCGUCGUCGUAGUU (SEQ ID5′ UUCCUUCUCGUCGUCGUAGdTdT (SEQ sense NO: 40) ID NO: 42) [964-982][964-982] siHsp47-4d human sense 5′ CUACGACGACGAGAAGGAAAAGCUG 5′ hsp47(SEQ ID NO: 43) CUACGACGACGAGAAGGAAAAGCdTdG [964-988] (SEQ ID NO: 45)[964-988] anti- 5′ 5′ sense CAGCUUUUCCUUCUCGUCGUCGUAGUUCAGCUUUUCCUUCUCGUCGUCGUAGUU (SEQ ID NO: 44) (SEQ ID NO: 46) [964-988][964-988] siHsp47-5 human sense 5′ GCCACACUGGGAUGAGAAAUU (SEQ5′ GCCACACUGGGAUGAGAAAdTdT (SEQ hsp47 ID NO: 47) ID NO: 49) [850-870][850-870] anti- 5′ UUUCUCAUCCCAGUGUGGCUU (SEQ ID5′ UUUCUCAUCCCAGUGUGGCdTdT (SEQ sense NO: 48) ID NO: 50) [850-868][850-868] siHsp47-6 human sense 5′ GCAGCAAGCAGCACUACAAUU (SEQ ID5′ GCAGCAAGCAGCACUACAAdTdT (SEQ hsp47 NO: 51) ID NO: 53) [675-693][675-693] anti- 5′ UUGUAGUGCUGCUUGCUGCUU (SEQ 5′ UUGUAGUGCUGCUUGCUGCdTdT(SEQ sense ID NO: 52) ID NO: 54) [675-693] [675-693] siHsp47-7 humansense 5′ CCGUGGGUGUCAUGAUGAUUU (SEQ 5′ CCGUGGGUGUCAUGAUGAUdTdT (SEQhsp47 ID NO: 55) ID NO: 57) [921-939] [921-939] anti-5′ AUCAUCAUGACACCCACGGUU (SEQ ID 5′ AUCAUCAUGACACCCACGGdTdT (SEQ senseNO: 56) ID NO: 58) [921-939] [921-939]

The contents of the articles, patents, and patent applications, and allother documents and electronically available information mentioned orcited herein, are hereby incorporated by reference in their entirety tothe same extent as if each individual publication was specifically andindividually indicated to be incorporated by reference.

Applicants reserve the right to physically incorporate into thisapplication any and all materials and information from any sucharticles, patents, patent applications, or other physical and electronicdocuments.

It will be readily apparent to one skilled in the art that varyingsubstitutions and modifications can be made to the invention disclosedherein without departing from the scope and spirit of the invention.Thus, such additional embodiments are within the scope of the presentinvention and the following claims. The present invention teaches oneskilled in the art to test various combinations and/or substitutions ofchemical modifications described herein toward generating nucleic acidconstructs with improved activity for mediating RNAi activity. Suchimproved activity can include improved stability, improvedbioavailability, and/or improved activation of cellular responsesmediating RNAi. Therefore, the specific embodiments described herein arenot limiting and one skilled in the art can readily appreciate thatspecific combinations of the modifications described herein can betested without undue experimentation toward identifying nucleic acidmolecules with improved RNAi activity.

The inventions illustratively described herein may suitably be practicedin the absence of any element or elements, limitation or limitations,not specifically disclosed herein. Thus, for example, the terms “a” and“an” and “the” and similar referents in the context of describing theinvention (especially in the context of the following claims) are to beconstrued to cover both the singular and the plural, unless otherwiseindicated herein or clearly contradicted by context. The terms“comprising”, “having,” “including,” containing”, etc. shall be readexpansively and without limitation (e.g., meaning “including, but notlimited to,”). Recitation of ranges of values herein are merely intendedto serve as a shorthand method of referring individually to eachseparate value falling within the range, unless otherwise indicatedherein, and each separate value is incorporated into the specificationas if it were individually recited herein. All methods described hereincan be performed in any suitable order unless otherwise indicated hereinor otherwise clearly contradicted by context. The use of any and allexamples, or exemplary language (e.g., “such as”) provided herein, isintended merely to better illuminate the invention and docs not pose alimitation on the scope of the invention unless otherwise claimed. Nolanguage in the specification should be construed as indicating anynon-claimed element as essential to the practice of the invention.Additionally, the terms and expressions employed herein have been usedas terms of description and not of limitation, and there is no intentionin the use of such terms and expressions of excluding any equivalents ofthe features shown and described or portions thereof, but it isrecognized that various modifications are possible within the scope ofthe invention claimed. Thus, it should be understood that although thepresent invention has been specifically disclosed by preferredembodiments and optional features, modification and variation of theinventions embodied therein herein disclosed may be resorted to by thoseskilled in the art, and that such modifications and variations areconsidered to be within the scope of this invention.

The invention has been described broadly and generically herein. Each ofthe narrower species and subgeneric groupings falling within the genericdisclosure also form part of the invention. This includes the genericdescription of the invention with a proviso or negative limitationremoving any subject matter from the genus, regardless of whether or notthe excised material is specifically recited herein. Other embodimentsarc within the following claims. In addition, where features or aspectsof the invention are described in terms of Markush groups, those skilledin the art will recognize that the invention is also thereby describedin terms of any individual member or subgroup of members of the Markushgroup.

1. The compound of claim 170, wherein the sense and antisense strandsare selected from the oligonucleotides described as SERPINH1_(—)2 (SEQID NOS: 60 and 127), SERPINH1_(—)6 (SEQ ID NOS: 63 and 130),SERPINH1_(—)45a (SEQ ID NOS: 98 and 165), SERPINH1_(—)51 (SEQ ID NOS:101 and 168) and SERPINH1_(—)51a (SEQ ID NOS: 105 and 172). 2-40.(canceled)
 41. A nucleic acid molecule, wherein: (a) the nucleic acidmolecule includes a sense strand and an antisense strand; (b) eachstrand of the nucleic acid molecule is independently 15 to 49nucleotides in length; (c) a 15 to 49 nucleotide sequence of theantisense strand is complementary to a sequence of an mRNA encodinghsp47; and (d) a 15 to 49 nucleotide sequence of the sense strand iscomplementary to the antisense strand and includes a 15 to 49 nucleotidesequence of an mRNA encoding hsp47.
 42. The nucleic acid molecule ofclaim 41, wherein said sequence of said antisense strand that iscomplementary to a sequence of an mRNA encoding human hsp47 comprises asequence complimentary to a sequence between nucleotides 600-800 of SEQID NO: 1, 801-899 of SEQ ID NO: 1, 900-1000 of SEQ ID NO: 1, or1001-1300 of SEQ ID NO:
 1. 43-55. (canceled)
 56. The nucleic acidmolecule of claim 41, wherein said antisense strand comprises a sequenceselected from the group consisting of SEQ ID NO: 4 or a portion thereof,SEQ ID NO: 6 or a portion thereof, SEQ ID NO: 8 or a portion thereof,SEQ ID NO: 10 or a portion thereof, SEQ ID NO: 12 or a portion thereof,SEQ ID NO: 14 or a portion thereof, SEQ ID NO: 16 or a portion thereof,SEQ ID NO: 18 or a portion thereof, SEQ ID NO: 20 or a portion thereof,SEQ ID NO: 22 or a portion thereof, SEQ ID NO: 24 or a portion thereof,SEQ ID NO: 26 or a portion thereof, SEQ ID NO: 28 or a portion thereof,SEQ ID NO: 30 or a portion thereof, SEQ ID NO: 32 or a portion thereof,SEQ ID NO: 34 or a portion thereof, SEQ ID NO: 36 or a portion thereof,SEQ ID NO: 38 or a portion thereof, SEQ ID NO: 40 or a portion thereof,SEQ ID NO: 42 or a portion thereof, SEQ ID NO: 44 or a portion thereof,SEQ ID NO: 46 or a portion thereof, SEQ ID NO: 48 or a portion thereof,SEQ ID NO: 50 or a portion thereof, SEQ ID NO: 52 or a portion thereof,SEQ ID NO: 54 or a portion thereof, SEQ ID NO: 56 or a portion thereof,and SEQ ID NO: 58 or a portion thereof.
 57. The nucleic acid molecule ofclaim 41, wherein said antisense strand comprises a sequence selectedfrom the group consisting of SEQ ID NO: 3 or a portion thereof, SEQ IDNO: 5 or a portion thereof, SEQ ID NO: 7 or a portion thereof, SEQ IDNO: 9 or a portion thereof, SEQ ID NO: 11 or a portion thereof, SEQ IDNO: 13 or a portion thereof, SEQ ID NO: 15 or a portion thereof, SEQ IDNO: 17 or a portion thereof, SEQ ID NO: 19 or a portion thereof, SEQ IDNO: 21 or a portion thereof, SEQ ID NO: 23 or a portion thereof, SEQ IDNO: 25 or a portion thereof, SEQ ID NO: 27 or a portion thereof, SEQ IDNO: 29 or a portion thereof, SEQ ID NO: 31 or a portion thereof, SEQ IDNO: 33 or a portion thereof, SEQ ID NO: 35 or a portion thereof, SEQ IDNO: 37 or a portion thereof, SEQ ID NO: 39 or a portion thereof, SEQ IDNO: 41 or a portion thereof, SEQ ID NO: 43 or a portion thereof, SEQ IDNO: 45 or a portion thereof, SEQ ID NO: 47 or a portion thereof, SEQ IDNO: 49 or a portion thereof, SEQ ID NO: 51 or a portion thereof, SEQ IDNO: 53 or a portion thereof, SEQ ID NO: 55 or a portion thereof, and SEQID NO: 57 or a portion thereof.
 58. The nucleic acid molecule of claim41, wherein the antisense strand and the sense strand are independently15-49 nucleotides in length. 59-72. (canceled)
 73. The nucleic acidmolecule of claim 41, wherein said antisense and sense strands areseparate polynucleotide strands. 74-75. (canceled)
 76. The nucleic acidmolecule of claim 41, wherein the sense and antisense strands are partof a single polynucleotide strand having both a sense and antisenseregion.
 77. (canceled)
 78. The nucleic acid molecule of claim 41,wherein the nucleic acid molecule is a double stranded molecule and hasa blunt end on both ends.
 79. The nucleic acid molecule of claim 41,wherein the nucleic acid molecule is a double stranded molecule and hasan overhang on both ends of the molecule. 80-85. (canceled)
 86. Thenucleic acid molecule of claim 41, wherein the nucleic acid molecule isa double stranded molecule and has a blunt end on one end of themolecule and an overhang on the other end of the molecule. 87-100.(canceled)
 101. The nucleic acid molecule of claim 41, wherein thenucleic acid molecule comprises one or more modifications or modifiednucleotides. 102-129. (canceled)
 130. A method for treating anindividual suffering from a disease associated with hsp47 comprisingadministering to said individual a nucleic acid molecule of claim 41 inan amount sufficient to reduce expression of hsp47.
 131. (canceled) 132.A composition comprising a nucleic acid molecule of claim 41 and apharmaceutically acceptable carrier. 133-139. (canceled)
 140. A doublestranded oligonucleotide compound having the structure (A1): (A1) 5′(N)x-Z 3′ (antisense strand) 3′ Z′—(N′)y-z″ 5′ (sense strand) whereineach of N and N′ is a nucleotide which may be unmodified or modified, oran unconventional moiety; wherein each of (N)x and (N′)y is anoligonucleotide in which each consecutive N or N′ is joined to the nextN or N′ by a covalent bond; wherein each of Z and Z′ is independentlypresent or absent, but if present independently includes 1-5 consecutivenucleotides or non-nucleotide moieties or a combination thereofcovalently attached at the 3′ terminus of the strand in which it ispresent; wherein z″ may be present or absent, but if present is acapping moiety covalently attached at the 5′ terminus of (N′)y; whereineach of x and y is independently an integer between 18 and 40; whereinthe sequence of (N′)y has complementary to the sequence of (N)x; andwherein (N)x includes an antisense sequence to SEQ ID NO:1.
 141. Thecompound of claim 140 wherein (N)x comprises an antisenseoligonucleotide present in Table A-19. 142-166. (canceled)
 167. A doublestranded oligonucleotide compound of claim 140 having a structure (A2)set forth below: (A2) 5′ N¹—(N)x-Z 3′ (antisense strand) 3′Z′—N²—(N′)y-z″ 5′ (sense strand) wherein each of N², N and N′ is anunmodified or modified ribonucleotide, or an unconventional moiety;wherein each of (N)x and (N′)y is an oligonucleotide in which eachconsecutive N or N′ is joined to the adjacent N or N′ by a covalentbond; wherein each of x and y is independently an integer between 17 and39; wherein the sequence of (N′)y has complementarity to the sequence of(N)x and (N)x has complementarity to a consecutive sequence in a targetRNA; wherein N¹ is covalently bound to (N)x and is mismatched to thetarget RNA or is a complementary DNA moiety to the target RNA; whereinN¹ is a moiety selected from the group consisting of natural or modifieduridine, deoxyribouridine, ribothymidine, deoxyribothymidine, adenosineor deoxyadenosine; wherein z″ may be present or absent, but if presentis a capping moiety covalently attached at the 5′ terminus of N²—(N′)y;and wherein each of Z and Z′ is independently present or absent, but ifpresent is independently 1-5 consecutive nucleotides, consecutivenon-nucleotide moieties or a combination thereof covalently attached atthe 3′ terminus of the strand in which it is present.
 168. The compoundof claim 167 wherein x=y=18.
 169. (canceled)
 170. The compound of claim167 wherein (N)x comprises an antisense oligonucleotide present in TableA-18. 171-172. (canceled)
 173. A composition comprising a compound ofclaim 140; and a pharmaceutically acceptable carrier.
 174. (canceled)175. The compound of claim 140 wherein at least one of Z or Z′ ispresent.
 176. The compound of claim 140 wherein the antisense strandcomprises 2′OMe sugar modified pyrimidines.
 177. A method for treatingan individual suffering from a disease associated with hsp47 comprisingadministering to said individual a nucleic acid molecule of claim 140 inan amount sufficient to reduce expression of hsp47.